TW202008302A - DNA-based data access by converting the input data into a set of nucleotide sequences and synthesizing a set of nucleic acids including the set of nucleotide sequences - Google Patents

Abstract

The present invention relates to DNA-based data storage. The present invention provides an exemplary method for storing the input data on a nucleic acid including: converting the input data into a set of nucleotide sequences and synthesizing a set of nucleic acids including the set of nucleotide sequences. The aforementioned conversion includes a data processing step and a nucleotide encoding step. The aforementioned data processing step includes converting the aforementioned input data into a binary string. The aforementioned nucleotide encoding step includes converting a binary string by using a 5-bit transcoding frame to obtain the aforementioned set of nucleotide sequences.

Description

DNA-based data access

The present invention generally relates to data storage and retrieval, and more specifically, to techniques for achieving reliable and effective DNA-based data storage and retrieval.

The use of DNA as a tool for data storage and retrieval (retrival) can be traced back to 1988, when Joe Davis and his collaborators created a synthetic DNA called "Mocrovenus" for coding icons (icon) And integrate it into E. coli cells. Compared with traditional storage media such as magnetic tapes and hard drives, DNA-based storage has a higher density (for example ~1mm ³ for storing 1EB data) and a longer storage period (for example, more than 1 million years at -18°C) And the advantage of lower maintenance costs. DNA storage is a prospective field of research based on oligonucleotide synthesis for DNA storage media generation (especially high-throughput synthesis platforms like CustomArray) and sequencing for information retrieval (especially next-generation sequencing (NGS), such as Illumina HiSeq 2500 and MiSeq).

However, at present, DNA-based data storage has many limitations. For example, the production cost of DNA synthesis is quite high, and the speed of data retrieval may be low due to sequencing. Therefore, DNA-based storage has been considered more suitable for large-scale file storage, which involves a smaller number of reads and writes to storage media. Further, many errors can be introduced at various stages of the process (such as encoding, writing, storing, decoding, reading, and retrieving), thereby jeopardizing the input and output of the data flow. Exemplary errors include mutations, deletions, insertions, losses, and denaturation after long-term storage of DNA fragments during synthesis and sequencing. In addition, when using DNA to store large amounts of data, it may be challenging to achieve random access to a portion of the data instead of fully retrieving the data.

The invention relates to a technology for realizing reliable and effective DNA-based data storage and retrieval. Specifically, the present invention provides an accurate, effective, and reliable method for storing input data on nucleic acids (eg, deoxyribonucleic acid ("DNA")). In particular, the present invention utilizes a novel 5-bit transcoding framework to convert one or more data files into nucleic acid sequences (eg, DNA sequences). The present invention further provides an integration process that includes a compression algorithm, an error correction algorithm, and a transcoding framework for efficient and reliable data storage and retrieval. In addition, the present invention allows random data access, which is particularly beneficial when storing large-scale data together, but only needs to browse part of the information at a given time. The data that can be stored according to the method of the present invention includes any type of data that can be represented digitally (ie, in the form of binary data), including, for example, text files, high-definition movies, images, and/or audio.

In some embodiments, a method for storing input data on a nucleic acid is provided. The method includes: a) converting the input data into a set of nucleotide sequences, wherein the conversion includes i) a data processing step, including converting the input data The data is converted into a binary string; and ii) a nucleotide coding step, including converting the binary string using a 5-bit transcoding framework to obtain a nucleotide sequence group; and b) synthesizing a nucleic acid group including the nucleotide sequence group .

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) data processing steps, including converting input data into a binary string; and ii) kernel The nucleotide coding step includes converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences.

In some embodiments, the data processing step includes dividing the binary string into a sequence of non-overlapping 5-bit binary strings.

In some embodiments, the nucleotide coding step includes converting each 5-bit binary string to an integer ranging from 0 to 31 to obtain an integer string.

In some embodiments, the nucleotide encoding step further includes converting the integer string using a 5-bit transcoding framework to obtain a set of nucleotide sequences.

In some embodiments, the nucleotide coding step further includes dividing the integer string into a plurality of initial integer subsequences with a predetermined length.

In some embodiments, the length of each of the plurality of initial integer subsequences is based on the oligomer length of the selected synthesis platform, the required error tolerance, the size of the input data, the selected error correction code or The combination is determined.

In some embodiments, the nucleotide coding step further includes adding index information to each of the plurality of initial integer subsequences to obtain a plurality of indexed integer subsequences.

In some implementations, the index information added to each of the plurality of initial integer sub-sequences includes an integer sequence, where the length of the integer sequence is based on the size of the input data.

In some embodiments, the nucleotide coding step includes, after adding the index information, adding redundant data to a plurality of indexed integer subsequences, thereby obtaining a plurality of redundant integer subsequences.

In some implementations, adding redundant data to multiple indexed integer subsequences includes: creating an empty matrix, where the number of columns in the empty matrix is greater than the size of multiple indexed integer subsequences, and The number of rows is greater than the number of integers in each of the multiple indexed integer subsequences; filling the empty matrix with multiple indexed integer subsequences and data generated by applying error correction coding; and obtaining based on the filled matrix Multiple integer subsequences with redundancy.

In some embodiments, the number of columns of the empty matrix is based on the length of the oligomer of the selected synthesis platform, the type of error correction code, the predetermined error tolerance value, the size of multiple indexed integer subsequences, or a combination thereof determine.

In some embodiments, the number of rows of the empty matrix is based on the oligomer length of the selected synthesis platform, the type of error correction code, the predetermined error tolerance value, the size of multiple indexed integer subsequences, or a combination thereof determine.

In some implementations, the error correction coding is Reed-Solomon ("RS") coding.

In some embodiments, the data generated by applying the error correction code is generated by applying RS code string correction and/or RS code block correction.

In some implementations, the 5-bit transcoding framework is based on Table 2.

In some embodiments, the selection of R and Y is based on: 1) a nucleotide different from the nucleotide immediately before R or Y; and/or 2) the estimated GC content of the nucleotide sequence.

In some implementations, the input data corresponds to compressed files. In some implementations, the input data corresponds to two or more files.

In some implementations, the input data corresponds to a text file.

In some implementations, the data processing further includes compressing the input data to obtain a compressed file and converting the compressed file into a binary string.

In some embodiments, the compressed file is compressed using the Lempel-Zic-Markov chain algorithm ("LZMA").

In some embodiments, the data processing step further includes: grouping two or more files into TAR files.

In some implementations, the TAR file is further compressed using the Lempel-Zic-Markov chain algorithm ("LZMA").

In some embodiments, the nucleotide coding step further includes appending primer sequence pairs to the 5'and 3'ends of each nucleotide sequence of the nucleotide sequence group.

In some embodiments, the primer pair is attached to the synthetic nucleic acid set.

In some embodiments, a method for storing two or more sets of input data on a nucleic acid is provided. The method includes: a) converting two or more sets of input data separately according to any method described in the present invention Into two or more sets of corresponding nucleotide sequences; b) appending primer sequence pairs to the 5'and 3'ends of each of the two or more sets of corresponding nucleotide sequences respectively, wherein The primer pairs of two or more sets of corresponding nucleotide sequences are different from each other; and c) Two or more sets of nucleic acids including the aforementioned two or more sets of corresponding nucleotide sequences are synthesized, respectively.

In some embodiments, each pair of primers has a sequence different from any one of two or more sets of corresponding nucleotide sequences or complementary sequences thereof.

In some embodiments, the GC content of the aforementioned synthetic nucleic acid group ranges from 30% to 70%. In some embodiments, the GC content of the aforementioned synthetic nucleic acid group ranges from less than about 70%.

In some embodiments, the aforementioned synthetic nucleic acid set is stored. In some embodiments, the aforementioned synthetic nucleic acid set is stored dry. In some embodiments, the aforementioned synthetic nucleic acid set is stored by lyophilization.

In some embodiments, the aforementioned synthetic nucleic acid set is fixed on a carrier. In some embodiments, the aforementioned carrier is a microarray.

In some embodiments, a method for retrieving output data stored on a nucleic acid is provided, the method includes: a) obtaining a nucleotide sequence group of a nucleic acid group, b) the nucleotide sequence The group is converted into output data, wherein the aforementioned conversion includes: i) a nucleotide decoding step, including converting the nucleotide sequence group into a binary string using a 5-bit transcoding framework; and ii) a data processing step, including the two The carry string is converted into output data to obtain the aforementioned output data.

In some embodiments, the nucleic acid set is amplified before the output data is retrieved.

In some embodiments, the nucleic acid set is sequenced to generate multiple sequence reads.

In some embodiments, multiple sequence reads are paired, merged, and filtered to obtain the aforementioned set of nucleotide sequences.

In some embodiments, a computer-implemented method for converting a set of nucleotide sequences into output data is provided. The method includes: i) a nucleotide decoding step, including using a 5-bit transcoding framework to convert the nucleotide The sequence group is converted into a binary string; and ii) the data processing step includes converting the binary string into output data.

In some embodiments, the nucleotide decoding step includes converting the set of nucleotide sequences into multiple integer subsequences including integers in the range of 0-31.

In some embodiments, the nucleotide decoding step further includes applying an error correction code to multiple integer subsequences to obtain multiple indexed integer subsequences.

In some embodiments, the step of applying error correction coding includes: i) applying RS coding string correction to multiple integer subsequences to obtain multiple uniform integer subsequences; and ii) applying RS coding block correction to multiple Consistent integer subsequences to obtain multiple indexed integer subsequences.

In some embodiments, the nucleotide decoding step further includes removing the index from the plurality of indexed integer subsequences to obtain a plurality of core integer subsequences.

In some embodiments, the nucleotide decoding step further includes combining core integer subsequences into integer strings.

In some embodiments, the nucleotide decoding step further includes converting the integer string to a binary string.

In some implementations, the output data is stored in a compressed file. In some embodiments, the data processing step further includes decompressing the compressed file. In some embodiments, the decompression is performed by LZMA algorithm.

In some implementations, the output data corresponds to multiple files. In some implementations, the aforementioned multiple files are extracted from the output data through the TAR algorithm.

In some implementations, the 5-bit transcoding framework is based on Table 2.

In some embodiments, the nucleic acid set includes primer sequences at the 5'and 3'ends and the aforementioned method includes removing the primer sequence before the nucleotide decoding step.

In some embodiments, a method for retrieving output data stored on a nucleic acid group of interest is provided, wherein the nucleic acid group of interest is one of multiple sets of nucleotide sequences present in the mixture, Each set encodes a different output data set and has different primer pair sets at the 3'and 5'ends. The method includes: a) amplifying the nucleic acid set using primer pairs corresponding to the nucleic acid of interest; b) obtained The nucleotide sequence group of the amplified nucleic acid, c) converting the nucleotide sequence group into output data according to the method in the above-mentioned embodiment mode; thereby obtaining the aforementioned output data.

In some embodiments, a method for retrieving corresponding two or more sets of output data stored on two or more sets of nucleic acids of interest is provided, wherein the aforementioned two or more sets of interest The nucleic acid is among multiple nucleotide sequences present in the mixture, each group encodes a different output data set and has different primer pair sets at the 3'and 5'ends. The method includes: a) using Primer pairs corresponding to the aforementioned two or more sets of nucleic acids of interest are amplified (eg, amplified separately or together) to the aforementioned two or more sets of nucleic acids of interest; b) two sets of amplified nucleic acids are obtained Or more sets of nucleotide sequences, c) according to any one of the methods described in the present invention, the two or more sets of nucleotide sequences are converted into two or more sets of output data; More sets of output data.

In some embodiments, a non-transitory computer-readable storage medium storing one or more programs is provided. The aforementioned one or more programs include instructions, which are executed by one or more processors of an electronic device When the electronic device is implemented, any method as described in the present invention.

The present invention further provides a system for providing nucleic acid-based data storage or retrieving data from nucleic acids, including: one or more processors; memory; and one or more programs, wherein one or more of the foregoing The program is stored in the memory and is configured to be executed by the aforementioned one or more processors. The aforementioned one or more programs include instructions for implementing any one of the methods described in the present invention.

The present invention further provides an electronic device for providing nucleic acid-based data storage or retrieving data from nucleic acids, the device comprising equipment for implementing any of the methods described in the present invention.

The present invention provides an accurate, effective, and reliable method for storing input data on nucleic acids, such as deoxyribonucleic acid ("DNA"). Specifically, the present invention utilizes a novel 5-bit transcoding framework to convert one or more data files into nucleic acid sequences (eg, DNA sequences). This new 5-bit transcoding framework allows efficient nucleic acid sequence design to strike the correct GC content and avoid certain homopolymers (such as homopolymers with 4 or more nucleotides in length), And reduce the error rate in nucleic acid synthesis and amplification. The present invention further provides an integration process that includes a compression algorithm, an error correction algorithm, and a transcoding framework for efficient and reliable data storage and retrieval. The method provided by the present invention can be used to store data of any size, including large-size files. In addition, the present invention allows random data access, which is particularly beneficial when storing large-scale data together, but only needs to browse part of the information at a given time. The data that can be stored according to the method of the present invention includes any type of data that can be represented digitally (ie, in the form of binary data), including, for example, text files, high-definition movies, images, and/or audio.

[Figure 1] Represents an exemplary process for providing DNA-based data storage and retrieval according to some implementations.

[FIG. 2] Represents exemplary means for processing compressed files for DNA-based data storage according to some implementations.

[FIG. 3A] Represents exemplary steps for adding indexes and redundant data to digital content to be stored according to some implementations.

[FIG. 3B] Depicts exemplary steps for adding indexes and redundant data to digital content to be stored according to some implementations.

[FIG. 3C] Depicts exemplary steps for adding indexes and redundant data to digital content to be stored according to some implementations.

[FIG. 3D] Depicts exemplary steps for adding indexes and redundant data to digital content to be stored according to some implementations.

[FIG. 4] Represents exemplary means for processing compressed files for DNA-based data storage according to some implementation types.

[Figure 5] Represents an exemplary 5-bit transcoding framework according to some implementations.

[FIG. 6] Represents an exemplary text portion to be stored and retrieved according to some implementations.

[FIG. 7] Represents an exemplary implementation of DNA-based data storage and retrieval technology according to some implementation types.

[FIG. 8] Depicts an exemplary electronic device according to some implementations.

[FIG. 9A] Represents an exemplary process for providing DNA-based data storage according to some implementations.

[FIG. 9B] Represents an exemplary process for providing DNA-based data retrieval according to some implementation types.

The present invention provides an accurate, effective, and reliable method for storing input data on nucleic acids, such as deoxyribonucleic acid ("DNA"). Specifically, the present invention utilizes a novel 5-bit transcoding framework to convert one or more data files into nucleic acid sequences (eg, DNA sequences). This new 5-bit transcoding framework allows efficient nucleic acid sequence design to strike the correct GC content and avoid certain homopolymers (such as homopolymers with 4 or more nucleotides in length), And reduce the error rate in nucleic acid synthesis and amplification. The present invention further provides an integration process that includes a compression algorithm, an error correction algorithm, and a transcoding framework for efficient and reliable data storage and retrieval. The method provided by the present invention can be used to store data of any size, including large-size files. In addition, the present invention allows random data access, which is particularly beneficial when storing large-scale data together, but only needs to browse part of the information at a given time. The data that can be stored according to the method of the present invention includes any type of data that can be represented digitally (ie, in the form of binary data), including, for example, text files, high-definition movies, images, and/or audio.

Therefore, in one aspect, the present invention provides a method for storing input data on a nucleic acid group and a method for converting the input data into a nucleotide sequence group. In another aspect, a method for retrieving output data stored on a nucleic acid and a method for converting a set of nucleotide sequences into output data are provided. Further provided is a system for storing one or more programs and a non-transitory computer-readable storage medium for implementing any one or more steps of the methods described in the present invention.

It should be understood that the embodiments of the present invention described in the present invention include "consisting of implementation forms" and/or "essentially consisting of implementation forms".

The reference to "about" a value or parameter in the present invention includes (and describes) a change to the value or parameter itself. For example, a description related to "about X" includes a description of "X".

As used in the present invention, the reference to a value or parameter that is "not" generally means and describes "except" the value or parameter. For example, this method is not used to treat type X cancer, meaning that the method is used to treat other types of cancer than X.

As used in the present invention and the appended patent application, the singular form includes plural indicators unless the context clearly dictates otherwise.

As used in the present invention and the appended patent applications, "a group" refers to one or more indicators unless the context clearly dictates otherwise. The nucleic acid group may be a nucleic acid encoding data of the same file or the same file compressed together. In some embodiments, the nucleic acids in the same file may have the same primer sets appended to the 5'and 3'ends.

Encoding data and data storage method

In one aspect, the present invention provides a method (for example, a computer-implemented method) for converting input data into a set of nucleotide sequences. The method generally includes a data processing step, which converts the input data into a binary string, and a nucleotide encoding step, which uses a 5-bit transcoding framework to convert the aforementioned binary string to obtain a set of nucleotide sequences. This method can be used to store input data on a nucleic acid group, which involves first converting the input data into a nucleotide sequence group, and then synthesizing the nucleic acid group including the aforementioned nucleotide sequence group.

The input data can represent any number of files of any type, such as text files, image files, video/audio files (such as high-definition files), etc. The file can be uncompressed or compressed. When the file is uncompressed, it can be compressed before being converted into a binary string. For example, the Lempel-Ziv-Markov Chain algorithm (Lempel-Ziv-Markov Chain algorithm) can be used to compress the files into LZMA files (eg A.lzma). In some embodiments, two or more files (such as three, four, five, six, and more files) are first grouped together, such as a TAR file (such as A.tar), And the TAR file is further compressed into an LZMA file (for example, A.tar.lzma). As such, the method can allow multiple files (eg, 1-5, 5-10, 10-15, 15-25, 25-35, 35-50) to be stored in a single nucleic acid composition.

In some implementations, random access to positions in a single file is allowed, the single file can be divided into multiple sets of data, and the multiple sets of data are each compressed and processed as described below. For example, a digitized file corresponding to a book with 10 chapters can be divided into 10 files, each file corresponding to a single chapter. Then compress and process the ten files to achieve free access to any chapter.

The data processing step converts the input data into a binary string. The binary string can be directly converted into a set of nucleotide sequences, for example, by following the 5-bit transcoding framework described in the present invention. Alternatively, the binary string can be further converted into an integer string, which is then converted into a set of nucleotide sequences, for example, by following a 5-bit transcoding framework. In some embodiments, the integer string is further subjected to error correction coding and/or other processing to generate multiple integer subsequences with redundancy, and then multiple integer subsequences with redundancy, for example, by following the 5-bit conversion The code frame is converted into a set of nucleotide sequences.

Therefore, for example, in some embodiments, a method (for example, a computer-implemented method) for converting input data into a set of nucleotide sequences is provided, wherein the conversion includes: i) a data processing step, including converting the input data Into a binary string; and ii) a nucleotide encoding step, which includes converting the aforementioned binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. In some embodiments, a method for storing input data on a nucleic acid is provided. The method includes: a) converting the input data into a set of nucleotide sequences, wherein the conversion includes i) a data processing step, including converting the input data The data is converted into a binary string; and ii) a nucleotide coding step, which includes converting the aforementioned binary string using a 5-bit transcoding framework to obtain a nucleotide sequence group; and b) synthesizing a nucleic acid group including the aforementioned nucleotide sequence group .

In some implementations, the data processing step includes dividing the binary string into a sequence of non-overlapping 5-bit binary strings, each of which can be further converted into integers ranging from integers 0 to 31 to obtain integer strings. This integer string can be directly converted into a set of nucleotide sequences, for example, using a 5-bit transcoding framework. Alternatively, the integer string is further operated as described below.

Specifically, the integer string may be divided into a plurality of initial integer sub-sequences having a predetermined length. The predetermined length of the initial integer subsequence is calculated based on a number of factors including the length of the oligomer of the synthesis platform, the selected error correction code, the required error tolerance, the synthesis error rate of the oligomer and/or the total The encoding data size is discussed in detail below. For example, the integer string can be divided into a column of non-overlapping integer subsequences using a sliding window of fixed length (eg, 22 integers). An index can then be added to each of the multiple initial integer subsequences to generate multiple indexed integer subsequences. The index may contain integers that are also in the range 0 to 31. The length of the index is flexible and depends on the yield of DNA synthesis and the size of the data.

In some embodiments, redundant data is added to generate multiple integer subsequences with redundancy. For example, Reed-Solomon (RS) error correction coding is applied to multiple integer subsequences to generate a new column of integer subsequences with redundancy through RS coded string correction and block correction. Redundancy refers to excess synthetic oligomers to provide robustness to dropout. Redundancy in string correction helps error correction of oligomer transitions and transversions. Redundancy in block correction can realize the correction of information insertion, deletion and complete loss.

In an exemplary implementation form, adding redundant data to multiple indexed integer subsequences includes: creating an empty matrix, where the number of columns in the empty matrix is greater than the size of multiple indexed integer subsequences, and wherein the empty The number of rows in the matrix is greater than the number of integers in each of multiple indexed integer subsequences; filling the empty matrix with multiple indexed integer subsequences and data generated by applying error correction coding; and based on the filled Obtains multiple integer subsequences with redundancy. The number of columns and/or rows of the empty matrix may be determined based on the type of error correction code, the predetermined error tolerance value, the size of multiple indexed integer subsequences, or a combination thereof. The error correction coding is Reed-Solomon ("RS") coding. In some embodiments, the data generated by applying the error correction code is generated by applying RS code string correction and RS code block correction.

In some embodiments, the nucleotide coding step further includes appending primer sequence pairs to the 5'and 3'ends of the nucleotide sequence set. The aforementioned primers can be used, for example, to amplify nucleic acid groups by PCR amplification methods. In some embodiments, the primer sequence is added to the set of nucleotide sequences before synthesis. Alternatively, the primer can be attached to the synthetic nucleic acid, for example, by ligation.

The aforementioned method can be used to store two or more sets of input data on a nucleic acid. Specifically, the method includes a) converting two or more sets of input data into two or more sets of corresponding nucleotide sequences; b) appending primer sequence pairs to the aforementioned two or more sets respectively The 5'and 3'ends of each of the nucleotide sequences of the above, wherein the primers of each of the aforementioned two or more corresponding sets of nucleotide sequences are different from each other, and c) the synthesis includes the aforementioned two or Multiple sets of nucleic acids with corresponding sets of nucleotide sequences. Each primer pair may have a sequence that is different from any one of two or more corresponding nucleotide sequences or complementary sequences thereof.

The synthetic nucleic acid may have a GC content of about 30% to about 70%. For example, the synthetic nucleic acid may have any of about 40% to about 60%, about 30% to about 40%, about 40% to about 50%, about 50% to about 60%, or about 60% to about 70% GC content. In some embodiments, the synthetic nucleic acid does not have a homopolymer longer than 3 nucleotides (eg, does not have a homopolymer of 4, 5, 6, 7, 8, 9, or 10 nucleotides). In some embodiments, the nucleic acid is an oligonucleotide, for example, an oligonucleotide of about 50, 150, 200, 300, or 400 nucleotides in length. In some embodiments, the nucleic acid set includes about 1, 2, 3, 5, 10, 15 or more oligonucleotides in any number.

In some embodiments, the aforementioned method further includes storing the synthetic nucleic acid set. In some embodiments, the nucleic acid set is stored by drying, for example, by lyophilization. The nucleic acid set can be stored as a dry composition, including a lyophilized composition. In some embodiments, the nucleic acid set is immobilized on a support, including a solid support such as a microarray. In some embodiments, nucleic acids are stored on a microarray with a density of about 5 μg per 1 inch×3 inch area (eg, in a CustomArray 12K wafer). In some implementations, the size of the input data is at least about 50MB.

Decoding nucleic acid sequence and data retrieval method

In another aspect, the present invention provides a method (for example, a computer-implemented method) for converting a set of nucleotide sequences into output data. This method is almost the opposite process of the coding program, and usually includes a nucleotide decoding step, which converts a nucleotide sequence group into a binary string, for example, by using a 5-bit transcoding framework, and a data processing step, which converts the binary Serial conversion into output data. The method can be used to retrieve output data stored on a nucleic acid group, which involves obtaining the nucleotide sequence of the nucleic acid group, and then converting the nucleotide sequence group into output data.

In some embodiments, the aforementioned nucleic acid group is first amplified, for example, by using primers present at the 3'and 5'ends of the nucleic acid group. And the amplified nucleic acid can be sequenced, such as next-generation sequencing. Next-generation sequencing technology is known to those of ordinary skill in the technical field. For example, nucleic acids are sequenced using the Illumina sequencing method. Sequences belonging to a specific file can be obtained by aligning primer sequences. In some embodiments, the method includes NGS library preparation. When the aforementioned nucleic acid group is present in a mixture including different nucleic acid groups encoding different materials, the nucleic acid group of interest can be specifically amplified by using the unique primer pair of the nucleic acid group of interest, thereby allowing the corresponding Random access to nucleic acid group data. If it is necessary to read and decode several compressed files in a single run of next-generation sequencing, then all their corresponding nucleic acid groups will be amplified by PCR and all corresponding pairs will be used.

In some embodiments, the method includes pair-end next-generation sequencing and read pairing and merging, where forward and reverse reads from a single cluster will be paired and merged into a single read, and all have New readings of regular length will be filtered. Furthermore, based on the primer sequence, all readings can be grouped for their respective compressed files. The primer can then be removed, and the nucleotide sequence can be converted into multiple integer subsequences including integers in the range of 0-31, or directly into a binary string, which is then converted into output data.

In some embodiments, the aforementioned method further includes applying error correction of multiple integer subsequences to obtain multiple indexed integer subsequences. In an exemplary embodiment, the step of applying error correction coding includes: i) applying RS coding string correction to multiple integer subsequences to obtain multiple uniform integer subsequences; and ii) applying RS coding block correction to The foregoing multiple uniform integer subsequences obtain multiple indexed integer subsequences. Because a nucleic acid can have many molecular copies and be sequenced multiple times during synthesis, many reads may represent a nucleic acid. These readings may change due to errors caused during high-throughput synthesis and sequencing, but correct readings that exactly match the originally designed nucleic acid still have a counting advantage. With the highest frequency-based correction at each position of the integer string, all integer strings sharing the same index can be corrected and merged into a consistent integer string between string correction and block correction.

Then indexes from multiple indexed integer subsequences can be removed to obtain multiple core integer subsequences. The integer string can then be concatenated into a complete integer string and then converted into a binary string. The binary string can then be written to a file, such as a compressed file. The compressed file can then be decompressed, for example, by using the LZMA algorithm. If the decompressed file includes data corresponding to multiple files, the decompressed file is further processed (eg, extracted) by the TAR algorithm to obtain the aforementioned multiple files.

In some embodiments, the aforementioned method can be used to retrieve the output data stored on the nucleic acid group of interest, where the nucleic acid group of interest is one of multiple sets of nucleotide sequences present in the mixture, each The sets encode different sets of output data and have different sets of primer pairs at the 3'and 5'ends. The method includes a) amplifying the aforementioned nucleic acid group using primer pairs corresponding to the nucleic acid group of interest; b) obtaining the nucleotide sequence group of the amplified nucleic acid group, c) and applying the core according to the method in the above-described embodiment mode The nucleotide sequence group is converted into output data; thereby obtaining the aforementioned output data.

In some embodiments, a method for retrieving corresponding two or more sets of output data stored on two or more sets of nucleic acids of interest is provided, wherein the aforementioned set of nucleic acids of interest is present in the mixture Among the multiple nucleotide sequence groups in each, each group encodes a different output data group and has different primer pair groups located at the 3'and 5'ends. The method includes: a) using corresponding to the aforementioned two groups or Primer pairs of more sets of nucleic acids of interest are amplified (eg, amplified separately or together) to the aforementioned two or more sets of nucleic acids of interest; b) two of the aforementioned two or more sets of amplified nucleic acids are obtained One or more sets of nucleotide sequences, and c) converting the aforementioned two or more sets of nucleotide sequences into two or more sets of output data, respectively; thereby obtaining the aforementioned two or more sets of output data.

5-bit transcoding framework

The method of the present invention utilizes a novel 5-bit transcoding framework for converting binary strings or integer strings into groups of nucleotide sequences. "5-bit transcoding framework" refers to the conversion according to Table 1 below. In general, every 5 consecutive bits from a binary string can be expressed as an integer between 0 and 31 and the following 3 nucleotides (ie, 3-mers). For example, nucleic acids have four bases (eg A, T, G and C), so the dimer (ie NN) should have 16 types (eg AA, AT, AG, AC, TA, TT, TG, TC, GA , GT, GG, GC, CA, CT, CG and CC). Assuming that the degenerate bases R and Y are connected after the dimer, the trimer (NNR/NNY) should consist of 32 species, which also matches well with the 32 integers in the range of 0 to 31 and makes the binary string Converted to DNA sequence.

In some embodiments, R is selected from any two of A, T, G, and C, and Y is selected from the other two of A, T, G, and C. In some embodiments, R is selected from A and G, and Y is selected from T and C. In some embodiments, R is selected from A and C, and Y is selected from T and G. In some embodiments, R is selected from T and G, and Y is selected from A and C. In some embodiments, R is selected from T and C, and Y is selected from A and G.

For example, for the purpose of maintaining the desired GC content and/or avoiding homopolymers, the choice of nucleotides corresponding to R and Y may depend on the base in front of them. For example, in one scheme R is selected from A and G and Y is selected from C and T, whether A or G is selected as R and whether C or T is selected as Y depends on the base in front of them (ie the second of the trimer Bases). In some embodiments, R and Y are selected so that the second and third bases are not the same. In some embodiments, R and Y are selected to maintain the desired GC balance. As long as the rules are followed, R and Y can be chosen randomly. The coding potential of this transcoding framework is 1.67 (ie 5 bits for 3nt).

Table 2 provides an exemplary 5-bit transcoding framework. In the specific embodiment depicted in Table 2, when Y is to be selected from C and Y, then R is selected from A and G. It will be understood that other transcoding frameworks that follow the same principle can be used.

Table 2

Synthesis and storage of nucleic acids

Nucleic acids including the desired nucleotide sequence can be synthesized using any nucleic acid synthesis method. In some embodiments, the nucleic acid is synthesized by chemical synthesis. The method of high-throughput nucleic acid synthesis is described in International Application No. WO 2002US40580 entitled "COMBINATORIAL SYNTHESIS ON ARRAYS" filed by Maurer et al. on February 17, 2002, and its publication number is WO 03052383 in December 2016. It is disclosed as "ELECTROCHEMICALLY GENERATED ACID AND ITS CONTAINMENT TO 100 MICRON REACTION AREAS FOR THE PRODUCTION OF DNA MICROARRAYS", which is incorporated by reference in its entirety.

Once synthesized, nucleic acids can be stored in different media. In some embodiments, nucleic acids are dried (eg, lyophilized) and stored in vials. In some embodiments, the nucleic acid is immobilized on a support, for example, a solid support such as a microarray.

Computer readable storage medium and system

The present invention further provides a non-transitory computer-readable storage medium that stores one or more programs. The aforementioned one or more programs include instructions that, when executed by one or more processors of an electronic device, cause the The electronic device implements one or more steps of any method described in the present invention.

In some embodiments, a system for providing nucleic acid-based data storage or retrieving data from a nucleic acid is provided, the system includes: one or more processors; memory; and one or more programs, Wherein the aforementioned one or more programs are stored in the memory and configured to be executed by the aforementioned one or more processors, the aforementioned one or more programs include one or more for implementing any of the methods described in the present invention Multi-step instructions.

In some embodiments, an electronic device for providing nucleic acid-based data storage or retrieving data from nucleic acids is provided. The device includes a device that implements any of the methods described in the present invention.

Exemplary implementation

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) data processing steps, including converting input data into a binary string; and ii) kernel The nucleotide coding step includes converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The data processing step includes dividing the binary string into a sequence of non-overlapping 5-bit binary strings. The nucleotide coding step includes converting each 5-bit binary string into an integer ranging from 0 to 31 to obtain an integer string, and converting the integer string using a 5-bit transcoding framework to obtain a nucleotide sequence group.

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) data processing steps, including converting the input data into a binary string; and ii) nucleotide encoding The steps include converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The data processing step includes dividing the binary string into a sequence of non-overlapping 5-bit binary strings. The nucleotide coding step includes converting each 5-bit binary string into an integer ranging from 0 to 31 to obtain an integer string, and converting the integer string using a 5-bit transcoding framework to obtain a nucleotide sequence group. The nucleotide coding step further includes dividing the integer string into a plurality of initial integer subsequences having a predetermined length.

In some embodiments, the length of each of the plurality of initial integer subsequences is based on the oligomer length of the selected synthesis platform, the required error tolerance, the size of the input data, the selected error correction code or The combination is determined.

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided: the method includes: i) data processing steps, including converting the input data into a binary string; and ii) nucleotides The coding step includes converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The data processing step includes dividing the binary string into a sequence of non-overlapping 5-bit binary strings. The nucleotide coding step includes converting each 5-bit binary string into an integer ranging from 0 to 31 to obtain an integer string, and converting the integer string using a 5-bit transcoding framework to obtain a nucleotide sequence group. The nucleotide coding step further includes dividing the integer string into a plurality of initial integer subsequences having a predetermined length. The nucleotide coding step further includes adding index information to each of the plurality of initial integer subsequences to obtain a plurality of indexed integer subsequences.

In some implementations, the index information added to each of the plurality of initial integer sub-sequences includes an integer sequence, where the length of the integer sequence is based on the size of the input data.

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) data processing steps, including converting input data into a binary string; and ii) kernel The nucleotide coding step includes converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The data processing step includes dividing the binary string into a sequence of non-overlapping 5-bit binary strings. The nucleotide coding step includes converting each 5-bit binary string into an integer ranging from 0 to 31 to obtain an integer string, and converting the integer string using a 5-bit transcoding framework to obtain a nucleotide sequence group. The nucleotide coding step further includes dividing the integer string into a plurality of initial integer subsequences having a predetermined length. The nucleotide coding step further includes adding index information to each of the plurality of initial integer subsequences to obtain a plurality of indexed integer subsequences. The nucleotide coding step further includes adding redundant data to multiple integer subsequences with indexes after adding index information, thereby obtaining multiple integer subsequences with redundancy.

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) data processing steps, including converting input data into a binary string; and ii) kernel The nucleotide coding step includes converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The data processing step includes dividing the binary string into a sequence of non-overlapping 5-bit binary strings. The nucleotide coding step includes converting each 5-bit binary string into an integer ranging from 0 to 31 to obtain an integer string, and converting the integer string using a 5-bit transcoding framework to obtain a nucleotide sequence group. The nucleotide coding step further includes dividing the integer string into a plurality of initial integer subsequences having a predetermined length. The nucleotide coding step further includes adding index information to each of the plurality of initial integer subsequences to obtain a plurality of indexed integer subsequences. The nucleotide coding step further includes adding redundant data to multiple integer subsequences with indexes after adding index information, thereby obtaining multiple integer subsequences with redundancy. Adding redundant data to multiple indexed integer subsequences includes: creating an empty matrix, where the number of columns in the empty matrix is greater than the size of multiple indexed integer subsequences, and where the number of rows in the empty matrix is greater than that in multiple The number of integers in each of the indexed integer subsequences; filling the empty matrix with multiple indexed integer subsequences and data generated by applying error correction coding; and obtaining multiple redundancy based on the filled matrix (Integer) subsequence.

In some embodiments, the number of empty matrix columns is determined based on the oligomer length of the selected synthesis platform, the type of error correction code, the predetermined error tolerance value, the size of multiple indexed integer subsequences, or a combination thereof .

In some embodiments, the number of rows of the empty matrix is determined based on the oligomer length of the selected synthesis platform, the type of error correction code, the predetermined error tolerance value, the size of multiple indexed integer subsequences, or a combination thereof .

In some implementations, the error correction code is Reed-Solomon ("RS") code.

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) data processing steps, including converting input data into a binary string; and ii) kernel The nucleotide coding step includes converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The data processing step includes dividing the binary string into a sequence of non-overlapping 5-bit binary strings. The nucleotide coding step includes converting each 5-bit binary string into an integer ranging from 0 to 31 to obtain an integer string, and converting the integer string using a 5-bit transcoding framework to obtain a nucleotide sequence group. The nucleotide coding step further includes dividing the integer string into a plurality of initial integer subsequences having a predetermined length. The nucleotide coding step further includes adding index information to each of the plurality of initial integer subsequences to obtain a plurality of indexed integer subsequences. The nucleotide coding step further includes adding redundant data to multiple integer subsequences with indexes after adding index information, thereby obtaining multiple integer subsequences with redundancy. Adding redundant data to multiple indexed integer subsequences includes: creating an empty matrix, where the number of columns in the empty matrix is greater than the size of multiple indexed integer subsequences, and where the number of rows in the empty matrix is greater than that in multiple The number of integers in each of the indexed integer subsequences; filling the empty matrix with multiple indexed integer subsequences and data generated by applying error correction coding; and obtaining multiple redundancy based on the filled matrix Integer subsequence. The material generated by applying error correction coding is generated by applying RS coded string correction and/or RS coded block correction.

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) converting the input data into a binary string; ii) dividing the binary string into non-overlapping The sequence of the 5-bit binary string; iii) convert each 5-bit binary string to an integer in the range of 0 to 31 to obtain an integer string and convert the integer string using a 5-bit transcoding framework; iv) convert the integer string Divided into multiple initial integer subsequences with a predetermined length; v) Add index information to each of the multiple initial integer subsequences to obtain multiple indexed integer subsequences; vi) After adding index information, add redundant data To multiple integer subsequences with indexes, to obtain multiple integer subsequences with redundancy, to obtain a nucleotide sequence group.

In some embodiments, a method for storing input data on a nucleic acid is provided. The method includes: i) converting the input data into a binary string; ii) dividing the binary string into non-overlapping 5-bit binary strings Iii) Convert each 5-bit binary string to an integer in the range of 0 to 31 to obtain an integer string and use a 5-bit transcoding framework to convert the integer string; iv) Divide the integer string into multiples with a predetermined length Initial integer sub-sequence; v) add index information to each of the initial integer sub-sequences to obtain multiple indexed integer sub-sequences; vi) after adding index information, add redundant data to multiple indexed integers Subsequences, thereby obtaining multiple integer subsequences with redundancy, thereby obtaining a nucleotide sequence group; and vii) synthesizing a nucleic acid group including the nucleotide sequence group.

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) converting the input data into a binary string; ii) dividing the binary string into non-overlapping Sequence of 5 bit binary strings; iii) convert each 5 bit binary string to an integer in the range of 0 to 31 to obtain an integer string and convert the integer string using a 5-bit transcoding framework; iv) convert the integer string Divided into multiple initial integer subsequences with a predetermined length; v) Add index information to each of the multiple initial integer subsequences to obtain multiple indexed integer subsequences; vi) Create an empty matrix, where the columns in the empty matrix The number is greater than the size of multiple indexed integer subsequences, and the number of rows in the empty matrix is greater than the number of integers in each of the multiple indexed integer subsequences; vii) use multiple indexed Integer subsequences and data generated by applying error correction coding to fill the empty matrix (for example, by applying RS-coded string correction and/or RS-coded block correction); and viii) obtaining multiple redundant data based on the filled matrix Integer subsequences to obtain a set of nucleotide sequences.

In some embodiments, a method for storing input data on a nucleic acid is provided. The method includes: i) converting the input data into a binary string; ii) dividing the binary string into non-overlapping 5-bit binary strings Iii) Convert each 5-bit binary string to an integer in the range of 0 to 31 to obtain an integer string and use a 5-bit transcoding framework to convert the integer string; iv) Divide the integer string into multiples with a predetermined length Initial integer sub-sequences; v) add index information to each of the multiple initial integer sub-sequences to obtain multiple indexed integer sub-sequences; vi) create an empty matrix, where the number of columns in the empty matrix is greater than multiple indexed The size of the integer subsequence of, and the number of rows in the empty matrix is greater than the number of integers in each of the multiple indexed integer subsequences; vii) use multiple indexed integer subsequences and pass the application The data generated by the error correction coding fills the empty matrix (for example, by applying RS-coded string correction and/or RS-coded block correction); and viii) obtaining multiple integer subsequences with redundancy based on the filled matrix, thereby obtaining The aforementioned nucleotide sequence group; and xi) synthesis of a nucleic acid group including the nucleotide sequence group.

In some embodiments, a method for retrieving output data stored on a nucleic acid is provided, the method comprising: i) obtaining a nucleotide sequence group of a nucleic acid group, ii) converting the nucleotide sequence group into Multiple integer subsequences including integers in the range of 0-31; iii) converting the nucleotide sequence group into a binary string; and iv) converting the binary string into output data, thereby obtaining the aforementioned output data.

In some embodiments, a method for retrieving output data stored on a nucleic acid is provided, the method comprising: i) sequencing a nucleic acid set to generate multiple sequence reads; ii) pairing, merging and/or filtering to obtain Nucleotide sequence group; iii) convert the nucleotide sequence group into multiple integer subsequences including integers in the range of 0-31; iv) apply error correction codes to the aforementioned multiple integer subsequences, thereby obtaining multiple Indexed integer subsequences; v) converting the aforementioned indexed integer subsequences into a binary string; and vi) converting the binary string into output data to obtain the aforementioned output data.

In some embodiments, a method for retrieving output data stored on a nucleic acid is provided, the method comprising: i) sequencing a nucleic acid set to generate multiple sequence reads; ii) pairing, merging and/or filtering to obtain Nucleotide sequence group; iii) convert the nucleotide sequence group into multiple integer subsequences including integers in the range of 0-31; iv) apply RS coding string correction to the foregoing multiple integer subsequences to obtain multiple Uniform integer subsequences; v) applying RS coding block correction to the foregoing multiple uniform integer subsequences to obtain multiple indexed integer subsequences; vi) converting the foregoing indexed integer subsequences to binary String; and vii) Convert the binary string into output data, thereby obtaining the aforementioned output data.

In some embodiments, a method for retrieving output data stored on a nucleic acid is provided, the method comprising: i) sequencing a nucleic acid set to generate multiple sequence reads; ii) pairing, merging and/or filtering to obtain Nucleotide sequence group; iii) convert the nucleotide sequence group into multiple integer subsequences including integers in the range of 0-31; iv) apply RS coding string correction to the foregoing multiple integer subsequences to obtain multiple Uniform integer subsequences; v) applying RS coding block correction to the foregoing multiple uniform integer subsequences to obtain multiple indexed integer subsequences; vi) removing indexes from the foregoing multiple indexed integer subsequences To obtain multiple core integer subsequences; vii) merge the aforementioned core integer subsequences into an integer string; viii) convert the aforementioned integer string into a binary string; and ix) convert the binary string into output data to obtain the aforementioned Output data.

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) data processing steps, including converting input data into a binary string; and ii) kernel The nucleotide coding step includes converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The aforementioned 5-bit transcoding framework is based on Table 2.

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) data processing steps, including converting input data into a binary string; and ii) kernel The nucleotide coding step includes converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The aforementioned 5-bit transcoding framework is based on Table 2. The selection of R and Y is based on: 1) different from the nucleotide immediately before R or Y; and/or 2) the estimated GC content of the nucleotide sequence.

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) data processing steps, including converting input data into a binary string; and ii) kernel The nucleotide coding step includes converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The input data corresponds to the compressed file. The compressed file is compressed using the Lempel-Zic-Markov chain algorithm ("LZMA").

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) data processing steps, including converting input data into a binary string; and ii) kernel The nucleotide coding step includes converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The input data corresponds to two or more files. The data processing step further includes: grouping two or more files into TAR files. The TAR file is further compressed using the Lempel-Zic-Markov chain algorithm ("LZMA").

In some embodiments, a computer-implemented method for converting input data into a set of nucleotide sequences is provided. The method includes: i) data processing steps, including converting input data into a binary string; and ii) kernel The nucleotide coding step includes converting the binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The nucleotide coding step further includes appending primer sequence pairs to the 5'and 3'ends of each nucleotide sequence of the aforementioned nucleotide sequence group.

In some embodiments, a method for storing input data on a nucleic acid includes a) converting the foregoing input data into a nucleotide sequence group, wherein the foregoing conversion includes i) a data processing step, including converting the input data into Binary string; ii) Nucleotide encoding step, including converting the binary string using a 5-bit transcoding framework to obtain a nucleotide sequence group; and b) Synthesizing a nucleic acid group including the nucleotide sequence group. The method further includes attaching a primer pair to the aforementioned synthetic nucleic acid set.

In some embodiments, a method for storing two or more sets of input data on a nucleic acid is provided. The method includes: a) according to any one of the methods described in the present invention, separate two or more sets of input data Convert into two or more sets of corresponding nucleotide sequences; b) Attach primer sequence pairs to the 5'and 3'ends of each of the two or more sets of corresponding nucleotide sequences, respectively, where The primer pairs of the aforementioned two or more sets of corresponding nucleotide sequences are different from each other; and c) Two or more sets of nucleic acids including the aforementioned two or more sets of corresponding nucleotide sequences are synthesized, respectively.

In some embodiments, a method for storing two or more sets of input data on a nucleic acid is provided. The method includes: a) according to any one of the methods described in the present invention, separate two or more sets of input data Convert into two or more sets of corresponding nucleotide sequences; b) Attach primer sequence pairs to the 5'and 3'ends of each of the two or more sets of corresponding nucleotide sequences, respectively, where The primer pairs of the aforementioned two or more sets of corresponding nucleotide sequences are different from each other; and c) Two or more sets of nucleic acids including the aforementioned two or more sets of corresponding nucleotide sequences are synthesized, respectively. Each pair of primers has a sequence different from any one of two or more sets of corresponding nucleotide sequences or complementary sequences thereof.

In some embodiments, the GC content of the synthetic nucleic acid group ranges from 30% to 70%.

In some embodiments, a method for storing input data on a nucleic acid is provided. The method includes a) converting the foregoing input data into a set of nucleotide sequences, wherein the foregoing conversion includes i) a data processing step, including converting the input The data is converted into a binary string; ii) a nucleotide coding step, which includes converting the binary string using a 5-bit transcoding framework to obtain a nucleotide sequence group; and b) synthesizing a nucleic acid group including the aforementioned nucleotide sequence group. The method further includes storing the aforementioned synthetic nucleic acid set.

In some embodiments, the aforementioned synthetic nucleic acid set is stored by drying. In some embodiments, the aforementioned synthetic nucleic acid set is stored by lyophilization.

In some embodiments, the synthetic nucleic acid set is fixed on a carrier, which may be a microarray.

In some embodiments, a method for retrieving output data stored on a nucleic acid is provided. The method includes: a) obtaining a nucleotide sequence group of a nucleic acid group, b) converting the nucleotide sequence group into Output data, wherein the aforementioned conversion includes: i) a nucleotide decoding step, including converting the nucleotide sequence group into a binary string using a 5-bit transcoding framework; and ii) a data processing step, including converting the binary string Output data to obtain the aforementioned output data. The method includes amplifying the aforementioned nucleic acid group before retrieving the output data.

In some embodiments, a method for retrieving output data stored on a nucleic acid is provided. The method includes: a) obtaining a nucleotide sequence group of a nucleic acid group, b) converting the nucleotide sequence group into Output data, wherein the aforementioned conversion includes: i) a nucleotide decoding step, including converting the nucleotide sequence group into a binary string using a 5-bit transcoding framework; and ii) a data processing step, including converting the binary string Output data to obtain the aforementioned output data. The method further includes sequencing the aforementioned nucleic acid set to generate multiple sequence reads. Multiple sequence reads are paired, combined and filtered to obtain the aforementioned set of nucleotide sequences.

In some embodiments, a computer-implemented method for converting a set of nucleotide sequences into output data is provided. The method includes: i) a nucleotide decoding step, including using a 5-bit transcoding framework to convert the nucleotide The sequence group is converted into a binary string; and ii) the data processing step includes converting the binary string into output data. The nucleotide decoding step converts the nucleotide sequence group into multiple integer subsequences including integers in the range of 0-31.

In some embodiments, a computer-implemented method for converting a set of nucleotide sequences into output data is provided. The method includes: i) a nucleotide decoding step, including using a 5-bit transcoding framework to convert the nucleotide The sequence group is converted into a binary string; and ii) the data processing step includes converting the binary string into output data. The nucleotide decoding step converts the nucleotide sequence group into multiple integer subsequences including integers in the range of 0-31. The nucleotide decoding step further includes applying an error correction code to the foregoing multiple integer subsequences, thereby obtaining multiple indexed integer subsequences.

In some embodiments, a computer-implemented method for converting a set of nucleotide sequences into output data is provided. The method includes: i) a nucleotide decoding step, including using a 5-bit transcoding framework to convert the nucleotide The sequence group is converted into a binary string; and ii) the data processing step includes converting the binary string into output data. The nucleotide decoding step converts the nucleotide sequence group into multiple integer subsequences including integers in the range of 0-31. The nucleotide decoding step further includes applying an error correction code to the foregoing multiple integer subsequences, thereby obtaining multiple indexed integer subsequences. The steps of applying error correction coding include: i) applying RS coding string correction to the foregoing multiple integer subsequences to obtain multiple uniform integer subsequences; and ii) applying RS coding block correction to the foregoing multiple uniform integer subsequences to Obtain multiple indexed integer subsequences.

In some embodiments, a computer-implemented method for converting a set of nucleotide sequences into output data is provided. The method includes: i) a nucleotide decoding step, including using a 5-bit transcoding framework to convert the nucleotide The sequence group is converted into a binary string; and ii) the data processing step includes converting the binary string into output data. The nucleotide decoding step converts the nucleotide sequence group into multiple integer subsequences including integers in the range of 0-31. The nucleotide decoding step further includes applying an error correction code to the foregoing multiple integer subsequences, thereby obtaining multiple indexed integer subsequences. The nucleotide decoding step further includes removing the index from the aforementioned plurality of indexed integer subsequences to obtain a plurality of core integer subsequences.

In some embodiments, a computer-implemented method for converting a set of nucleotide sequences into output data is provided. The method includes: i) a nucleotide decoding step, including using a 5-bit transcoding framework to convert the nucleotide The sequence group is converted into a binary string; and ii) the data processing step includes converting the binary string into output data. Store the output data in a compressed file. The data processing step further includes, for example, by decompressing the compressed file through the LZMA algorithm.

In some embodiments, a computer-implemented method for converting a set of nucleotide sequences into output data is provided. The method includes: i) a nucleotide decoding step, including using a 5-bit transcoding framework to convert the nucleotide The sequence group is converted into a binary string; and ii) the data processing step includes converting the binary string into output data. The output data corresponds to multiple files. The method further includes extracting the aforementioned multiple files from the output data through the TAR algorithm.

In some embodiments, a computer-implemented method for converting a set of nucleotide sequences into output data is provided. The method includes: i) a nucleotide decoding step, including using a 5-bit transcoding framework to convert the nucleotide The sequence group is converted into a binary string; and ii) the data processing step includes converting the binary string into output data. The nucleotide decoding step converts the nucleotide sequence group into multiple integer subsequences including integers in the range of 0-31. The nucleotide decoding step further includes applying an error correction code to the aforementioned multiple integer subsequences, thereby obtaining multiple indexed integer subsequences. The nucleotide decoding step further includes removing the index from the aforementioned plurality of indexed integer subsequences to obtain a plurality of core integer subsequences. The nucleotide decoding step further includes merging the core integer subsequences into an integer string and converting the aforementioned integer string into a binary string.

In some embodiments, a computer-implemented method for converting a set of nucleotide sequences into output data is provided. The method includes: i) a nucleotide decoding step, including using a 5-bit transcoding framework to convert the nucleotide The sequence group is converted into a binary string; and ii) the data processing step includes converting the binary string into output data. The aforementioned 5-bit transcoding framework is based on Table 2.

In some embodiments, a computer-implemented method for converting a set of nucleotide sequences into output data is provided. The method includes: i) a nucleotide decoding step, including using a 5-bit transcoding framework to convert the nucleotide The sequence group is converted into a binary string; and ii) the data processing step includes converting the binary string into output data. The aforementioned nucleic acid set includes primer sequences at the 5'and 3'ends and the method includes removing the aforementioned primer sequence before the nucleotide decoding step.

In some embodiments, a computer feasible method for DNA-based data storage is provided. The method includes: converting a digitized file into a binary string; using a 5-bit transcoding framework to convert the aforementioned binary string to obtain an integer string ; Obtain multiple integer subsequences from the aforementioned integer string; and convert the foregoing multiple integer subsequences into multiple DNA oligomers for the synthesis of DNA.

In some embodiments, using the 5-bit transcoding framework to convert the aforementioned binary string to obtain an integer string includes: dividing the binary string into a sequence of non-overlapping 5-bit binary strings; converting each 5-bit binary string into Integers in the range 0 to 31 to obtain integer strings. In some embodiments, the aforementioned integer string is further divided into a plurality of initial integer sub-sequences having a predetermined length. In some implementations, obtaining multiple integer subsequences to be converted includes: adding index information to each of the initial multiple integer subsequences; after adding index information, adding redundant data to the initial multiple integers Subsequence to obtain multiple integer subsequences. In some implementations, the index information added to each initial plurality of subsequences includes integer strings, and the length of the integer string corresponding to the index information is based on the size of the digitized file.

In some embodiments, the foregoing method includes adding redundant data to multiple integer subsequences, which may include, for example, obtaining a subset of the initial multiple integer subsequences; selecting an empty matrix, where the number of columns in the empty matrix is greater than the sub The number of subsequences in the set, and the number of rows in the empty matrix is greater than the number of integers in each subsequence of the subset; fill the empty matrix with a subset of the initial multiple integer subsequences and data corresponding to the error correction code; And obtain multiple integer subsequences based on the filled matrix. In some implementations, the number of columns of the empty matrix is selected based on the type of error correction code, the predetermined error tolerance value, the size of the subset, or a combination thereof. In some implementations, the number of rows of the empty matrix is selected based on the type of error correction code, the predetermined error tolerance value, the size of the subset, or a combination thereof.

In some implementations, the error correction code is a Reed-Solomon ("RS") code. In some embodiments, the conversion of multiple integer subsequences into multiple DNA oligomers includes the conversion of the integers of the foregoing multiple integer subsequences into three nucleotides, where: these three The first of the nucleotides is selected from A, T, G and C, the second of these three nucleotides is selected from A, T, G and C, and the third of these three nucleotides is selected from One of two options.

In some embodiments, the digitized file is a compressed file corresponding to the group consisting of one or more files or directories. In some implementations, the digitized files include LZMA files corresponding to the group consisting of one or more files or directories compressed using the Lampel-Zico-Markov chain algorithm.

In some implementation forms according to any one of the implementation forms above, wherein the aforementioned method further comprises: adding data representing the primer pair to each oligomer expression form of the plurality of DNA oligomer expression forms; After the correct information, DNA synthesis is performed based on the representation of multiple DNA oligomers.

In some embodiments, the foregoing method further includes: obtaining a second digitized file; obtaining a representation of the second plurality of DNA oligomers based on the second digitized file; adding data representing the second primer pair to the second plurality of DNA Each oligomer representation of the oligomer, where the second primer pair is different from the first primer pair; and based on the representation of multiple DNA oligomers and the representation of second multiple DNA oligomers Perform DNA synthesis.

In some embodiments, a computer-feasible method for DNA-based data retrieval is provided. The method includes: obtaining multiple readings corresponding to digitized files; based on the foregoing multiple readings, obtaining multiple integer subsequences; Converting the aforementioned multiple integer subsequences into an integer string; using a 5-bit frame to convert the aforementioned integer string into a binary string; and obtaining a digitized file based on the binary string. In some embodiments, obtaining multiple readings corresponding to the aforementioned digital file includes identifying primers pre-associated with the digital file. In some embodiments, obtaining multiple integer subsequences includes performing frequency-based error correction based on multiple readings. In some implementations, using a 5-bit transcoding framework to convert an integer string to a binary string includes: converting each integer of the integer string to a 5-bit binary number.

In some embodiments, a non-transitory computer-readable storage medium storing one or more programs is provided. The aforementioned one or more programs include instructions, which are executed by one or more processors of an electronic device When the electronic device: convert the digitized file into a binary string; use a 5-bit transcoding framework to convert the aforementioned binary string to obtain an integer string; obtain multiple integer subsequences from the foregoing integer string; and convert the multiple integer substrings The sequence is converted into a representation of multiple DNA oligomers for DNA synthesis.

In some embodiments, a system for providing DNA-based data storage is provided. The foregoing system includes: one or more processors; a memory; and one or more programs, wherein the aforementioned one or more programs Stored in memory and configured to be executed by one or more processors, the aforementioned one or more programs include: instructions for converting digitized files into binary strings; for converting the aforementioned two using a 5-bit encoding framework Instructions for obtaining a string of integers; instructions for obtaining multiple integer subsequences from the aforementioned integer string; and instructions for converting the aforementioned multiple integer subsequences into representations of multiple DNA oligomers.

In some embodiments, a non-transitory computer-readable storage medium storing one or more programs is provided. The aforementioned one or more programs include instructions, which are executed by one or more processors of an electronic device When the electronic device obtains multiple readings corresponding to the digitized file; based on the multiple readings, obtains multiple integer subsequences; converts the multiple integer subsequences into integer strings; uses a 5-bit frame to convert the foregoing integer strings into Binary string; and obtaining a digitized file based on the aforementioned binary string.

In some embodiments, a system for providing DNA-based data storage is provided. The foregoing system includes: one or more processors; a memory; and one or more programs, wherein the aforementioned one or more programs Stored in the memory and configured to be executed by the aforementioned one or more processors, the aforementioned one or more programs include: instructions for obtaining a plurality of readings corresponding to the digitized file; for obtaining based on the aforementioned plurality of readings Instructions for multiple integer subsequences; instructions for converting the aforementioned multiple integer subsequences into integer strings; instructions for converting the aforementioned integer strings into binary strings using a 5-bit frame; and for use based on the aforementioned binary strings Get instructions for digital files.

According to an exemplary implementation method, the different steps of the aforementioned method are implemented by one or more computer software programs, which include software instructions designed to be executed by the data processor of the relay module according to the invention and designed to control the method Software instructions for the execution of different steps.

Therefore, on the one hand, the present invention also relates to a program that is easy to be executed by a computer or by a data processor. This program includes commands to control the execution of the steps of the aforementioned method.

This method can use any programming language in the form of source code, object code, or code between source code and object code, for example, in the form of partial compilation or in any other desired form.

The invention also relates to an information medium that can be read by the data processor and includes instructions of the program as described above.

The information medium may be any entity or device capable of storing programs. For example, the medium may include a storage device such as a ROM (which stands for "read only memory"), such as a CD-ROM (which stands for "optical disk read only memory") or a microelectronic circuit ROM or magnetic recording device, such as a floppy disk Or hard drive.

Further, the information medium may be a carrier that can be transmitted by radio or other means, for example, an electrical signal or an optical signal that can be delivered through a cable and an optical cable. In particular, the program can be downloaded into an Internet-type network.

Alternatively, the information medium may be an integrated circuit containing the aforementioned program, which is suitable for performing or for performing the method in question.

According to an implementation form, the implementation form of the present invention is implemented by means of software and/or hardware components. From this point of view, the term "module" in this document may correspond to software components and hardware components or a group of hardware and software components.

A software component corresponds to one or more computer programs, a subprogram of one or more programs, or more generally to any element of a program or a software program, which can implement one according to the content described below for the modules involved Function or set of functions. Such a software component is executed by a data processor of a physical entity (terminal, server, etc.) and can access the physical entity (memory, recording medium, communication bus, input/output electronic board, user interface, etc.) Hardware resources.

Similarly, a hardware element corresponds to any element of a hardware unit that can implement a function or a group of functions according to the content described below for the involved module. It can be a programmable hardware component or a component with an integrated circuit for executing software, such as an integrated circuit, a smart card, a memory card, an electronic board for executing firmware, etc. In a variant, the hardware element includes a processor as an integrated circuit, such as a central processing unit and/or a microprocessor and/or a dedicated integrated circuit (ASIC) and/or a dedicated instruction set processor (ASIP) and/or Or graphics processing unit (GPU) and/or physical processing unit (PPU) and/or digital signal processor (DSP) and/or image processor and/or auxiliary processor and/or floating point unit and/or network Processor and/or audio processor and/or multi-core processor. In addition, the hardware component may also include a baseband processor (including, for example, a memory unit and firmware) and/or a radio electronic circuit (which may include wires) that receives or transmits radio signals. In one embodiment, the hardware components comply with one or more standards, such as ISO/IEC 18092/ECMA-340, ISO/IEC 21481/ECMA-352, GSMA, StoLPaN, ETSI/SCP (Smart Card Platform), GlobalPlatform (Ie secure element). In a variant, the hardware element is a radio frequency identification (RFID) tag. In one embodiment, the hardware components include circuits that implement Bluetooth communication and/or Wi-Fi communication and/or Zigbee communication and/or USB communication and/or FireWire communication and/or NFC (for near field) communication.

It should be noted that the step of obtaining the element/value in the present invention may be regarded as a step of reading such element/value in the memory unit of the electronic device or receiving such element/value from another electronic device through communication means A step of.

Exemplary process

FIG. 1 shows an exemplary process for providing DNA-based data storage and retrieval according to some embodiments. Specifically, exemplary steps 102-110 involve encoding digital data for storage, and exemplary steps 112-122 involve decoding stored information for retrieval. Hereinafter, referring to FIGS. 2-5, the exemplary steps in FIG. 1 will be described in further detail.

1. Coding

In step 102 ("data compression"), one or more files and/or directories are packaged into a single file, and then compressed into a compressed file. In some embodiments, the file and/or directory is packaged into a TAR file (for example, File.tar), and then the Lampere-Zico-Malkov Chain Algorithm (ie, LZMA Algorithm) Compressed into LZMA file (for example, File.tar.lzma). In some embodiments, one LZMA file operates as a single non-splitter unit for data retrieval (eg, during decoding). Therefore, if multiple files and directories are to be stored together but retrieved randomly and independently, they should be grouped into multiple TAR files and compressed into multiple corresponding LZMA files at this step.

In step 104, the first round of data transcoding is implemented. First, convert each LZMA file to a binary string. As an example, referring to FIG. 2, a file named "File.tar.lzma" is converted to a binary string. Then convert the binary character string into an integer string B ("0; 10; 25; ...; 4; 8; 31"). In the depicted embodiment, the conversion from a binary string to an integer string B is achieved using a 5-bit transcoding framework. As shown in the figure, the binary string is divided into a series of non-overlapping 5-bit binary strings, such as "00000" and "01010". Each 5-bit binary string is then converted to an integer to form an integer string B. Those of ordinary skill in the art should know that, under the 5-bit transcoding framework, each integer in the integer string ranges from 0 (corresponding to "00000") to 31 (corresponding to "11111").

As shown in Fig. 2, a fixed-length sliding window is then used to divide the integer string B into multiple non-overlapping integer subsequences (eg, [A1, A2, ..., An]). In the embodiment depicted in FIG. 2, as depicted in FIG. 2, each integer subsequence (eg, A1) consists of 22 integers. Finally, the index information is appended to the beginning of each subsequence to form a new multiple integer subsequence with indexes (eg, [B1, B2, ..., Bn]). In the depicted embodiment, the index information includes a sequence of 3 integers, each integer ranging from 0 to 31. The length of the index sequence can be selected based on various factors, such as the size of the compressed file and the yield of DNA synthesis.

Returning to FIG. 1, in step 106, multiple indexed integer subsequences (for example, [B1, B2, ..., Bn] as shown in FIG. 2) are further transformed into multiple indexed and redundant The remaining integer subsequences (for example, [C1, C2, ..., Cm] as shown in FIG. 4). Various error correction coding algorithms, such as Reed-Solomon (RS) coding, fountain coding, and hamming coding, can be used to add redundant data to the digital data to be stored. In a preferred embodiment, RS coding is used because of its robustness and ease of implementation.

Figures 3A-D show how to add indexes and redundancy to digital content (eg, represented by multiple integer subsequences [A1, A2, ..., An]) to obtain [C1, C2, ..., Cm ]'S exemplary process. Specifically, FIGS. 3A-D show how to use RS encoding to process the first five integer subsequences (ie, A1, A2, A3, A4, and A5) to form [C1, C2, ..., C31]. For the remaining integer sub-sequences (ie, A6,...An), every five consecutive integer sub-sequences are treated as a unit in a manner similar to that shown in FIGS. 3A-D. In this embodiment, the five integer subsequences are processed together via a 29×31 matrix, so that the parity of the block correction is 26 (ie, 31-5=26), so 13 of 31 (ie, 26/ 2=13) oligomers may be lost, but can be recovered according to the principle of RS encoding.

Referring to FIG. 3A, prepare a 29×31 empty matrix, and fill the matrix with the first five integer strings A1, A2, A3, A4, A5 from [A1, A2, ..., An], which are shown to occupy 22× Sub-matrix of 5. This area is the central data block.

Returning to FIG. 3B, an index sequence consisting of three integers ranging from 0 to 31 is appended to the beginning of each column as a unique index, and the index string can be stored before appending. As shown in the figure, the index will be stored or allocated in ascending order, such as 0-0-0, 0-0-1, 0-0-2, ..., 0-0-31, 0-1-31, ... . In FIG. 3B, the indexed integer strings are labeled B1, B2, B3, B4, and B5, respectively.

Referring to FIG. 3C, RS coding is used to fill the blank area of each line occupied by the core data block line by line. This step is called "block correction" and helps to deal with, for example, lost oligomers and insertion deletions (including insertions and deletions) and long-term storage denaturation during synthesis and sequencing.

Returning to FIG. 3D, RS coding is used to fill the blank area of each column of the entire matrix column by column. This step is called "string correction" and helps correct for point mutations caused during synthesis, sequencing, and long-term storage, for example. As shown in FIG. 3D, the matrix now includes 31 integer strings [C1, C2, ..., C31]. In other words, after the block correction and the string correction, the aforementioned 5 integer sub-sequences A1-A5 are converted into 31 integer sub-sequences C1-C31. In addition, each of A1-A5 contains 22 integers, while each of C1-C31 contains 29 integers (including 3 additional index integers and 4 parity extra integers for RS encoding for error correction) . It should be understood that the various dimensions shown in FIGS. 3A-D are merely exemplary. The length of the index string (3 in Figures 3A-D), the size of the matrix (for example, 29×31 in Figures 3A-D), and the number of integer strings to be treated as units (for example, 5 in Figures 3A-D) ) Can be selected based on a variety of factors, such as the type of error code used, the required fault tolerance, and the characteristics of the DNA synthesis platform.

As shown in FIG. 4, through a round of string correction of RS coding and a round of block correction of RS coding according to the technique described with reference to FIGS. 3A-D, multiple integer subsequences with indexes, [B1, B2, ..., Bn], converted to multiple integer subsequences with redundancy, [C1, C2, ..., Cm], where m is greater than n. In addition, each integer in the integer subsequence [C1, C2, ..., Cm] ranges from 0 to 31.

In the embodiment depicted in Figures 3A-D. The length of the initial integer sub-sequence such as A1 (22 in the depicted embodiment) is calculated based on multiple factors. Specifically, the length of the integer string with index and redundancy (denoted as L, 29 in the depicted embodiment) is calculated from the oligomer length of the synthesis platform. The parity check and block correction of the two strings (denoted as X, 4 in the depicted embodiment) are determined by the synthetic error rate of the oligomer, the error correction code used, and the required error tolerance rate. The index length (denoted as Y, 3 in the depicted embodiment) is determined by the total coded data size. Therefore, the length of the initial integer string (expressed as Z) is Z=L-X-Y.

Returning to FIG. 1, in step 108, a second round of transcoding is implemented to convert the list of redundant integer strings (eg, [C1, C2, ..., Cm]) into a representation of multiple DNA oligomers (For example, [D1, D2, ..., Dm]). Each form of DNA oligomer contains the bases A, T, G, and C used for synthesis. In particular, the "5-bit transcoding framework" can be used again. Here, each integer in the integer string [C1, C2, ..., Cm] ranges from 0 to 31, so it can be uniquely mapped to one of 32 kinds of 3 nucleotides (for example, trimer, Including NNY and NNR, where N represents A, T, G, C; Y represents C and T; and R represents A and G). For example, as shown in Figure 5, the integer 6 corresponds to the 5-bit binary string "00110" and can be translated into "AGR" under a specific strategy. In some embodiments, the 5-bit transcoding framework can provide a direct conversion between integers and representations of DNA oligomers without any intermediate steps (eg, first convert the integer to a binary string).

Thus, each of the 29 integers (eg, C1) in each integer subsequence can be mapped to 3 nucleotides. After converting all [C1, C2, ..., Cm], replace Y with C or T, and replace R with A or G before DNA synthesis. This is done to ensure that the third base is different from the second base of the trimer, and to avoid three consecutive identical bases (eg, AAA, GGG, TTT, CCC). In addition, by choosing Y and R, the GC percentage of each oligomer should be limited to 30% to 70%. The replacement step not only reduces the error caused by oligomer synthesis, but also has important significance for improving the correction ratio of oligomer synthesis.

According to the principle of RS coding, tolerable errors may include two from each oligomer of the same matrix in the exemplary scheme shown in FIGS. 3A-D (ie, string-corrected parity check, 4 Half) mutations and 13 out of 31 oligomers (ie, block corrected parity, half of 26) lost oligomers (including completely lost oligomers and oligomers with indels).

Referring to FIG. 1, in step 110, a primer pair is added and DNA synthesis is performed. In some embodiments, a single compressed file (eg, File.tar.lzma of FIG. 4) is converted into multiple representations of DNA oligomers (eg, [D1, D2, ..., Dn in FIG. 4 ]). And the expression form of adding the same primer sequence pair at the two ends of each oligomer corresponding to the compressed file. For multiple compression files to be stored and synthesized simultaneously but requiring random access during subsequent reading and decoding, a unique orthogonal primer pair is selected for and associated with each compression file. For example, if there are 3 compressed files to be stored and synthesized at the same time but need to be randomly accessed during subsequent reading and decoding, then select 3 pairs of unique orthogonal primers to be associated with each of the 3 compressed files. For each compressed file, the selected primer pair is appended to each oligomer in the multiple oligomers corresponding to the compressed file. Then, all the oligomers corresponding to the aforementioned multiple compression files can be combined and synthesized together into a storage medium at the same time.

To select primer pairs, a variety of criteria can be used. For example, primer pairs can be selected to avoid homodimers, heterodimers, hairpin structures and have sufficient specificity (eg, there is no binding site for the encoding nucleic acid sequence). In some examples, multiple PCR primer design standards are used.

2. Decoding

The decoding program is basically the reverse process of the encoding program. Referring to FIG. 1, in step 112, PCR is performed using primer pairs to amplify the corresponding compressed file (for example, File.tar of FIG. 4) oligomer list (for example, [D1, D2, ..., in FIG. 4 Dn]). If you need to read and decode multiple compressed files with a single run of NGS, you should amplify all their corresponding oligomer lists by PCR using all corresponding primer pairs. This step is also called "NGS library preparation".

In step 114, double-ended next-generation sequencing and read pairing and merging are performed (eg, by Illumina sequencing system). Specifically, the forward and reverse readings from the same cluster are paired and merged into a single reading, and all new readings with irregular lengths (eg, readings with insertion deletions) will be filtered. In addition, based on the primer sequence, all readings can be grouped for each compressed file. In subsequent steps, the readings corresponding to the same compressed file (ie, readings sharing the same primer) will be analyzed together.

In step 116, reverse RS coding is performed. In some embodiments, a 29 by 31 zero matrix but a non-empty matrix will be used. Specifically, each reading from a single compressed file has PCR primers removed at both ends, which are then converted into integer subsequences by RS-encoded string correction with the purpose of error correction for mutations. Because an oligomer may have many molecular copies and be sequenced multiple times during synthesis, many of the above reads may be derived from an oligomer. These readings may change due to errors caused during high-throughput synthesis and sequencing, but correct readings should dominate. Through the highest frequency-based correction at each position of the integer subsequence, all integer subsequences sharing the same index can be corrected and merged into a uniform integer subsequence. For example, for a group of readings that share the same index, each position of a consistent integer subsequence should be determined by the integer that occurs most frequently at that position.

At step 118, the list of integer strings can be fully decoded by RS-encoded block correction, recovering lost oligomers and oligomers with insertions and deletions. Since an oligomer may have many molecular copies and be sequenced multiple times during synthesis, many readings may represent an oligomer. These readings may change due to errors caused during high-throughput synthesis and sequencing, but correct readings that are well matched to the originally designed oligomers still have a counting advantage. Through the highest frequency-based correction at each position of the integer string, all integer strings sharing the same index can be corrected and merged into a consistent integer string between word string correction and block correction. Since oligomers with insertions and deletions have irregular lengths and will be deleted during error correction, the corresponding data is completely equal to lack of information and needs to be restored. Based on the index information, the columns of the matrix are filled in after the correction based on the highest frequency.

In step 120, transcoding is performed. Readings are stored by index, and then the index is deleted from each integer subsequence. Then all integer sub-sequences can be connected into a single integer string, which is then transferred into a binary string through a 5-bit transcoding framework.

In step 122, decompression is performed. Specifically, the system writes the binary string to the compressed file, and then decompresses the compressed file through the LZMA algorithm and the TAR algorithm in sequence. For random access to multiple compressed files, steps 116 to 122 should be performed independently for each compressed file. The pool can store multiple compressed files. Each compressed file has its own PCR primer. During decoding, it is not necessary to sequence the entire pool. Instead, the corresponding PCR primers are used to amplify the oligomer of a certain compressed file, and then sequence the amplified oligomer to decode this corresponding compressed file rather than the entire pool.

As discussed above, a 5-bit transcoding framework is utilized. Specifically, every 5 consecutive bits from the binary string can be expressed as an integer between 0 and 31 and the following 3 nucleotides [nt] (ie, trimer). For example, DNA oligomers are composed of four bases (eg, A, T, G, and C), so there should be 16 dimers (ie, NN) (eg, AA, AT, AG, AC, TA, TT, TG, TC, GA, GT, GG, GC, CA, CT, CG and CC). Assuming that the degenerate bases R and Y are connected after the dimer, the trimer (NNR/NNY) should consist of 32 species, which also matches well with 32 integers in the range of 0 to 31 and makes the binary string good To the DNA sequence. In the process of oligomer synthesis, whether to choose A or G to represent R and whether to choose C or T to replace Y depends on the base in front of them (ie the second base of the trimer). In fact, the aforementioned system can Make the second and third bases different, and at the same time maintain GC balance. In view of this premise, the exact bases will be randomly selected between candidate bases. In short, the coding potential of the conversion framework is 1.67 (ie 5 bits for 3nt).

FIG. 7 shows an exemplary implementation of DNA-based data storage and retrieval technology. Here, a text file (data size: 1.16 kb) containing Chinese characters as shown in FIG. 6 is stored via DNA according to the process described in the present invention.

During encoding, the text file is compressed into a single compressed file, and then 403 oligomers with a length of 87 nt are stored through a DNA storage framework. At the same time, in order to simulate random access, 6 copies of the compressed file are used and 6 pairs of primers are selected. Each pair of primers was added to both ends of each of the 403 oligomers. The aforementioned 6 pairs of primers (each 20 nt) are orthogonal, which means that any two of them have a sufficient Hamming distance and have less similarity to any of the 403 oligomers. The sequence table in the ASCII text file submitted here includes SEQ ID NO.1-SEQ ID NO.403 and primer pairs PP NO.1-PP NO.6 as SEQ ID NO.404-415.

Then the synthesis of the oligomer pool is carried out. A total of 2418 (ie 403 times 6) oligomers were synthesized using the CustomArray platform developed by CustomArray, Inc. Each oligomer is 127 nt, which includes a total of 40 nt primers (20 nt at each end).

Then PCR amplification and NGS are performed. Perform 6 PCR reactions on all compressed file copies. After using TruSeq DNA PCR-free HT library preparation kit (96 indexes in plate format, 96 samples) and 6 library indexes to prepare 6 samples, due to the length of oligomer 127nt, use MiSeq kit V3 (150 cycles) sequenced pooled samples together. The Q30 of NGS data is 94% (official standard>85%), and the cluster density is 1,301K/mm ² (official standard 1200-1400K/mm ² ).

Finally, decode. After each copy of the compressed file is independently decoded, all copies can be retrieved and decompressed randomly without any errors.

Figure 8 presents an apparatus that can be used to perform one or more steps of the method of the present invention. This device labeled 800 includes a computing unit (eg, "Central Processing Unit" CPU), labeled 801, and one or more memory units (eg, RAM ("Random Access Memory") blocks (where the The result can be a temporary storage during the execution of the instructions of the computer program), or a ROM block that stores the computer program, among other things, or an EEPROM ("electronically erasable and rewritable read-only memory") block or flash memory block ), marked as 802. The computer program can be composed of instructions executed by the computing unit. Such a device 800 may also include a dedicated unit labeled 803, which constitutes an input-output interface to allow the device 800 to communicate with other devices. In particular, this dedicated unit 803 can be connected to an antenna (for communication without contact), or to a serial port (for communication "contact"). It should be noted that these units can exchange data together via, for example, a bus.

In alternative implementations, some or all of the steps of the previously described method can be in a programmable FPGA ("Field Programmable Gate Array") component or an ASIC ("dedicated integrated circuit" ) Implemented in the hardware of the component.

In alternative embodiments, some or all of the steps of the previously described method may be performed on an electronic device that includes a memory unit and a processing unit (as disclosed in FIG. 8). This device 800 can be used in combination with a high-throughput synthesis platform (eg CustomArray) and a DNA sequencer (eg MiSeq sequencer).

9A depicts an exemplary method 900 for storing input data on nucleic acids. At block 902, the input data is converted into a set of nucleotide sequences. At block 904, the input data is converted into a binary string. At block 906, the binary string is converted using a 5-bit transcoding framework to obtain a set of nucleotide sequences. At block 908, a nucleic acid group including the nucleotide sequence group is synthesized.

9B depicts an exemplary method 950 for retrieving output data stored on nucleic acids. At block 952, a nucleotide sequence set of nucleic acid sets is obtained. At block 954, the set of nucleotide sequences is converted into output data. Specifically, at block 956, the set of nucleotide sequences is converted into a binary string using a 5-bit transcoding framework. At block 958, the binary string is converted into output data.

Although the present invention and embodiments have been fully described with reference to the accompanying drawings, it should be noted that various changes and modifications will be apparent to those of ordinary skill in the art. These changes and modifications should be understood to be included within the scope of the contents and embodiments disclosed by the scope of the patent application.

For the purpose of explanation, the foregoing description has been described with reference to specific embodiments. However, the above illustrative discussion is not intended to be exhaustive or to limit the invention to the precise forms disclosed. In light of the above teachings, many modifications and changes are possible. The selected and described embodiments are to best explain the principle of the technology and its practical application. Therefore, those of ordinary skill in the art can best utilize various modified techniques and various implementations suitable for the specific intended use.

This application requires the rights and interests of China Patent Application No. 201710611123.2 filed on July 25, 2017, the entire contents of which are incorporated by reference into the present invention for all purposes.

The following content submitted in an ASCII text file is incorporated into the present invention by reference in its entirety: Computer readable form (CRF) of sequence table (file name: application number 107127162 sequence table electronic data. TXT, record date: November 30, 2018 , Size: 179KB).

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<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 57

<210> 58

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 58

<210> 59

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 59

<210> 60

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 60

<210> 61

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 61

<210> 62

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 62

<210> 63

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 63

<210> 64

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 64

<210> 65

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 65

<210> 66

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 66

<210> 67

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 67

<210> 68

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 68

<210> 69

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 69

<210> 70

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 70

<210> 71

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 71

<210> 72

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 72

<210> 73

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 73

<210> 74

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 74

<210> 75

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 75

<210> 76

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 76

<210> 77

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 77

<210> 78

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 78

<210> 79

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 79

<210> 80

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 80

<210> 81

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 81

<210> 82

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 82

<210> 83

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 83

<210> 84

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 84

<210> 85

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 85

<210> 86

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 86

<210> 87

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 87

<210> 88

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 88

<210> 89

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 89

<210> 90

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 90

<210> 91

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 91

<210> 92

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 92

<210> 93

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 93

<210> 94

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 94

<210> 95

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 95

<210> 96

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 96

<210> 97

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 97

<210> 98

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 98

<210> 99

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 99

<210> 100

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 100

<210> 101

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 101

<210> 102

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 102

<210> 103

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 103

<210> 104

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 104

<210> 105

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 105

<210> 106

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 106

<210> 107

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 107

<210> 108

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 108

<210> 109

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 109

<210> 110

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 110

<210> 111

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 111

<210> 112

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 112

<210> 113

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 113

<210> 114

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 114

<210> 115

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 115

<210> 116

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 116

<210> 117

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 117

<210> 118

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 118

<210> 119

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 119

<210> 120

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 120

<210> 121

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 121

<210> 122

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 122

<210> 123

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 123

<210> 124

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 124

<210> 125

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 125

<210> 126

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 126

<210> 127

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 127

<210> 128

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 128

<210> 129

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 129

<210> 130

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 130

<210> 131

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 131

<210> 132

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 132

<210> 133

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 133

<210> 134

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 134

<210> 135

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 135

<210> 136

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 136

<210> 137

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 137

<210> 138

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 138

<210> 139

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 139

<210> 140

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 140

<210> 141

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 141

<210> 142

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 142

<210> 143

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 143

<210> 144

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 144

<210> 145

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 145

<210> 146

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 146

<210> 147

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 147

<210> 148

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 148

<210> 149

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 149

<210> 150

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 150

<210> 151

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 151

<210> 152

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 152

<210> 153

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 153

<210> 154

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 154

<210> 155

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 155

<210> 156

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 156

<210> 157

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 157

<210> 158

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 158

<210> 159

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 159

<210> 160

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 160

<210> 161

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 161

<210> 162

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 162

<210> 163

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 163

<210> 164

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 164

<210> 165

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 165

<210> 166

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 166

<210> 167

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 167

<210> 168

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 168

<210> 169

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 169

<210> 170

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 170

<210> 171

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 171

<210> 172

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 172

<210> 173

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 173

<210> 174

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 174

<210> 175

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 175

<210> 176

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 176

<210> 177

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 177

<210> 178

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 178

<210> 179

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 179

<210> 180

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 180

<210> 181

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 181

<210> 182

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 182

<210> 183

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 183

<210> 184

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 184

<210> 185

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 185

<210> 186

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 186

<210> 187

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 187

<210> 188

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 188

<210> 189

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 189

<210> 190

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 190

<210> 191

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 191

<210> 192

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 192

<210> 193

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 193

<210> 194

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 194

<210> 195

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 195

<210> 196

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 196

<210> 197

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 197

<210> 198

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 198

<210> 199

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 199

<210> 200

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 200

<210> 201

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 201

<210> 202

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 202

<210> 203

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 203

<210> 204

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 204

<210> 205

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 205

<210> 206

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 206

<210> 207

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 207

<210> 208

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 208

<210> 209

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 209

<210> 210

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 210

<210> 211

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 211

<210> 212

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 212

<210> 213

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 213

<210> 214

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 214

<210> 215

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 215

<210> 216

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 216

<210> 217

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 217

<210> 218

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 218

<210> 219

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 219

<210> 220

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 220

<210> 221

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 221

<210> 222

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 222

<210> 223

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 223

<210> 224

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 224

<210> 225

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 225

<210> 226

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 226

<210> 227

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 227

<210> 228

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 228

<210> 229

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 229

<210> 230

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 230

<210> 231

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 231

<210> 232

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 232

<210> 233

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 233

<210> 234

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 234

<210> 235

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 235

<210> 236

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 236

<210> 237

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 237

<210> 238

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 238

<210> 239

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 239

<210> 240

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 240

<210> 241

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 241

<210> 242

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 242

<210> 243

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 243

<210> 244

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 244

<210> 245

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 245

<210> 246

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 246

<210> 247

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 247

<210> 248

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 248

<210> 249

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 249

<210> 250

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 250

<210> 251

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 251

<210> 252

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 252

<210> 253

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 253

<210> 254

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 254

<210> 255

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 255

<210> 256

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 256

<210> 257

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 257

<210> 258

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 258

<210> 259

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 259

<210> 260

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 260

<210> 261

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 261

<210> 262

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 262

<210> 263

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 263

<210> 264

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 264

<210> 265

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 265

<210> 266

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 266

<210> 267

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 267

<210> 268

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 268

<210> 269

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 269

<210> 270

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<222> Synthetic construct

<400> 270

<210> 271

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 271

<210> 272

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 272

<210> 273

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 273

<210> 274

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 274

<210> 275

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 275

<210> 276

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 276

<210> 277

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 277

<210> 278

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 278

<210> 279

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 279

<210> 280

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 280

<210> 281

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 281

<210> 282

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 282

<210> 283

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 283

<210> 284

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 284

<210> 285

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 285

<210> 286

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 286

<210> 287

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 287

<210> 288

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 288

<210> 289

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 289

<210> 290

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 290

<210> 291

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 291

<210> 292

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 292

<210> 293

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 293

<210> 294

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 294

<210> 295

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 295

<210> 296

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 296

<210> 297

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 297

<210> 298

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 298

<210> 299

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 299

<210> 300

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 300

<210> 301

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 301

<210> 302

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 302

<210> 303

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 303

<210> 304

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 304

<210> 305

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 305

<210> 306

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 306

<210> 307

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 307

<210> 308

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 308

<210> 309

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 309

<210> 310

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 310

<210> 311

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 311

<210> 312

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 312

<210> 313

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 313

<210> 314

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 314

<210> 315

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 315

<210> 316

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 316

<210> 317

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 317

<210> 318

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 318

<210> 319

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 319

<210> 320

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 320

<210> 321

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 321

<210> 322

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 322

<210> 323

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 323

<210> 324

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 324

<210> 325

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 325

<210> 326

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 326

<210> 327

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 327

<210> 328

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 328

<210> 329

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 329

<210> 330

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 330

<210> 331

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 331

<210> 332

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 332

<210> 333

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 333

<210> 334

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 334

<210> 335

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 335

<210> 336

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 336

<210> 337

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 337

<210> 338

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 338

<210> 339

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 339

<210> 340

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 340

<210> 341

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 341

<210> 342

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 342

<210> 343

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 343

<210> 344

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 344

<210> 345

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 345

<210> 346

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 346

<210> 347

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 347

<210> 348

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 348

<210> 349

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 349

<210> 350

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 350

<210> 351

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 351

<210> 352

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 352

<210> 353

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 353

<210> 354

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 354

<210> 355

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 355

<210> 356

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 356

<210> 357

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 357

<210> 358

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 358

<210> 359

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 359

<210> 360

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 360

<210> 361

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 361

<210> 362

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 362

<210> 363

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 363

<210> 364

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 364

<210> 365

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 365

<210> 366

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 366

<210> 367

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 367

<210> 368

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 368

<210> 369

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 369

<210> 370

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 370

<210> 371

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 371

<210> 372

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 372

<210> 373

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 373

<210> 374

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 374

<210> 375

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 375

<210> 376

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 376

<210> 377

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 377

<210> 378

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 378

<210> 379

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 379

<210> 380

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 380

<210> 381

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 381

<210> 382

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 382

<210> 383

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 383

<210> 384

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 384

<210> 385

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 385

<210> 386

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 386

<210> 387

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 387

<210> 388

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 388

<210> 389

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 389

<210> 390

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 390

<210> 391

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 391

<210> 392

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 392

<210> 393

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 393

<210> 394

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 394

<210> 395

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 395

<210> 396

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 396

<210> 397

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 397

<210> 398

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 398

<210> 399

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 399

<210> 400

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 400

<210> 401

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 401

<210> 402

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 402

<210> 403

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 403

<210> 404

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 404

<210> 405

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 405

<210> 406

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 406

<210> 407

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 407

<210> 408

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 408

<210> 409

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 409

<210> 410

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 410

<210> 411

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 411

<210> 412

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 412

<210> 413

<211> 20

<212> DNA

<213> Artificial sequence

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<223> Synthetic construct

<400> 413

<210> 414

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 414

<210> 415

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic construct

<400> 415

Claims (58)

A method for storing input data on a nucleic acid, characterized in that it includes: a) converting the foregoing input data into a nucleotide sequence group, wherein the foregoing conversion includes i) a data processing step, which includes converting the foregoing input data into two The carry string; ii) a nucleotide coding step, which includes converting the aforementioned binary string using a 5-bit transcoding framework to obtain the aforementioned nucleotide sequence group; and b) synthesizing the nucleic acid group including the aforementioned nucleotide sequence group. A computer-implemented method for converting input data into a set of nucleotide sequences, characterized in that it includes: i) a data processing step, which includes converting the aforementioned input data into a binary string; ii) a nucleotide encoding step, which This involves converting the aforementioned binary string using a 5-bit transcoding framework to obtain a set of nucleotide sequences. The method as described in item 1 or 2 of the patent application scope, wherein the aforementioned data processing step includes dividing the aforementioned binary string into a sequence of non-overlapping 5-bit binary strings. The method as described in item 3 of the patent application scope, wherein the aforementioned nucleotide coding step includes converting each 5-bit binary string into an integer ranging from 0 to 31 to obtain an integer string. The method as described in item 4 of the patent application scope, wherein the nucleotide encoding step further includes converting the integer string with the 5-bit transcoding framework to obtain the nucleotide sequence group. The method as described in item 4 of the patent application scope, wherein the aforementioned nucleotide coding step further includes dividing the aforementioned integer string into a plurality of initial integer subsequences having a predetermined length. The method as described in item 6 of the patent application scope, wherein the length of each of the plurality of initial integer subsequences is based on the length of the oligomer of the selected synthesis platform, the required error tolerance, and the size of the input data 3. Determine the selected error correction code or a combination thereof. The method as described in item 6 or 7 of the patent application scope, wherein the nucleotide encoding step further includes adding index information to each of the plurality of initial integer subsequences to obtain a plurality of indexed integer subsequences. The method as recited in item 8 of the patent application scope, wherein the index information added to each of the plurality of initial integer subsequences includes an integer sequence, wherein the length of the integer sequence is based on the size of the input data. The method as described in item 8 or 9 of the patent application, wherein the nucleotide coding step includes, after adding the index information, adding redundant data to the multiple indexed integer subsequences, thereby obtaining multiple With redundant integer subsequences. The method as described in item 10 of the patent application scope, wherein adding redundant data to the aforementioned multiple indexed integer subsequences includes: creating an empty matrix, wherein the number of columns in the aforementioned empty matrix is greater than the aforementioned multiple indexed integers The size of the subsequence, and wherein the number of rows in the empty matrix is greater than the number of integers in each of the plurality of indexed integer subsequences; encoding with the plurality of indexed integer subsequences and by applying error correction The generated data fills the aforementioned empty matrix; and obtains the aforementioned multiple integer subsequences with redundancy based on the filled matrix. The method as described in item 11 of the patent application range, wherein the number of columns of the empty matrix is based on the length of the oligomer of the selected synthesis platform, the type of error correction code, the predetermined error tolerance value, the plurality of indexed The size of the integer subsequence or a combination thereof is determined. The method as described in item 11 or 12 of the patent application scope, wherein the number of rows of the aforementioned empty matrix is based on the length of the oligomer of the selected synthesis platform, the type of error correction code, the predetermined error tolerance value, the plurality of The size of the indexed integer subsequence or a combination thereof is determined. The method as described in any of items 11-13 of the patent application scope, wherein the aforementioned error correction code is a Reed-Solomon ("RS") code. The method described in item 14 of the patent application scope, wherein the data generated by applying the error correction coding is generated by applying the string correction of the RS coding and/or the block correction of the RS coding. The method as described in any of items 1-15 of the patent application scope, wherein the aforementioned 5-bit transcoding framework is based on Table 2. The method as described in item 16 of the patent application scope, wherein the selection of R and Y is based on: 1) different from the nucleotide immediately before R or Y; and/or 2) the estimated GC content of the aforementioned nucleotide sequence . The method as described in any of items 1-17 of the patent application scope, wherein the aforementioned input data corresponds to a compressed file. The method as described in any of items 1 to 18 of the patent application scope, wherein the aforementioned input data corresponds to two or more files. The method as described in any of items 1-17 or 19 of the patent application scope, wherein the aforementioned input data corresponds to a text file. The method as described in any one of items 1-20 of the patent application scope, wherein the aforementioned data processing step further includes compressing the aforementioned input data to obtain a compressed file and converting the aforementioned compressed file into a binary string. The method as described in item 18 or 21 of the patent application scope, wherein the aforementioned compression file is compressed using the Lampel-Zico-Marykov Chain Algorithm ("LZMA"). As described in Item 19 of the patent application scope, the aforementioned data processing step further includes: grouping two or more files into TAR files. The method as described in item 23 of the patent application scope, in which the aforementioned TAR file is further compressed using the Lampel-Zico-Markov Chain Algorithm ("LZMA"). The method as described in any one of claims 1-24, wherein the aforementioned nucleotide coding step further includes appending a primer sequence pair to the 5'of each nucleotide sequence of the aforementioned nucleotide sequence group And the 3'end. The method as described in item 1 of the patent application scope, which further includes attaching a primer pair to the synthetic nucleic acid group. A method for storing two or more sets of input data on a nucleic acid, the characteristics of which include: a) As described in any one of items 2-19 of the patent application scope, input the two or more sets of the foregoing Convert the data into two or more sets of corresponding nucleotide sequences; b) Attach the primer sequence pairs to the 5'and 3'of each of the two or more sets of corresponding nucleotide sequences respectively Ends, where the primer pairs used for the aforementioned two or more sets of corresponding nucleotide sequences are different from each other; and c) synthesize two or more sets including the aforementioned two or more sets of corresponding nucleotide sequences, respectively Nucleic acid. The method as described in item 27 of the patent application range, wherein each pair of primers has a sequence different from any one of the aforementioned two or more sets of corresponding nucleotide sequences or complementary sequences thereof. The method as described in any of items 1 or 3-28 of the patent application scope, wherein the GC content of the synthetic nucleic acid group ranges from 30% to 70%. The method as described in any of items 1 or 3-29 of the patent application scope, wherein the GC content of the synthetic nucleic acid group is less than about 70%. The method as described in item 1 of the patent application scope, which further includes storing the synthetic nucleic acid group. The method as described in item 31 of the patent application scope, wherein the aforementioned synthetic nucleic acid group is stored by drying. The method as described in item 32 of the patent application scope, wherein the aforementioned synthetic nucleic acid group is stored by lyophilization. The method as described in item 31 of the patent application scope, wherein the aforementioned synthetic nucleic acid group is fixed on a carrier. The method described in item 34 of the patent application scope, wherein the aforementioned carrier is a microarray. A method for retrieving output data stored on a nucleic acid, characterized by: a) obtaining a nucleotide sequence group of a nucleic acid group, b) converting the aforementioned nucleotide sequence group into the aforementioned output data, wherein the aforementioned conversion Including: i) nucleic acid decoding step, including converting the aforementioned nucleotide sequence group into a binary string using a 5-bit transcoding framework; and ii) data processing step, including converting the binary string into the aforementioned output data, thereby obtaining the aforementioned output data. The method as described in item 36 of the patent application scope, wherein the foregoing method includes amplifying the nucleic acid group before retrieving the output data. The method as described in any one of the patent application items 36-37, further comprising sequencing the aforementioned nucleic acid group to generate multiple sequence reads. The method as described in item 38 of the patent application scope, wherein the plurality of sequence reads are paired, combined and filtered to obtain the aforementioned nucleotide sequence group. A computer-implemented method for converting a nucleotide sequence group into output data, which includes: i) a nucleotide decoding step, including using a 5-bit transcoding framework to convert the aforementioned nucleotide sequence group into a binary string ; And ii) data processing steps, including converting the binary string into the aforementioned output data. The method as described in any one of patent application items 36-40, wherein the nucleotide decoding step includes converting the nucleotide sequence group into multiple integer subsequences including integers in the range of 0-31 . The method as described in item 41 of the patent application range, wherein the nucleotide decoding step further includes applying an error correction code to the plurality of integer subsequences, thereby obtaining the plurality of indexed integer subsequences. The method as described in item 42 of the patent application range, wherein the step of applying error correction coding includes: i) applying RS code string correction to the foregoing multiple integer subsequences to obtain multiple uniform integer subsequences; and ii) The RS coding block correction is applied to the aforementioned plurality of uniform integer subsequences to obtain the aforementioned plurality of indexed integer subsequences. The method as recited in item 42 or 43 of the patent application range, wherein the nucleotide decoding step further includes removing the index from the plurality of indexed integer subsequences to obtain a plurality of core integer subsequences. The method as described in item 44 of the patent application range, wherein the aforementioned nucleotide decoding step further includes combining the aforementioned core integer subsequences into an integer string. The method as described in item 45 of the patent application scope, wherein the nucleotide decoding step further includes converting the integer string into a binary string. The method as described in item 46 of the patent application scope, in which the aforementioned output data is stored in a compressed file. The method as described in item 47 of the patent application scope, wherein the aforementioned data processing step further includes decompressing the aforementioned compressed file. The method as described in item 48 of the patent application scope, wherein the decompression is performed through the LZMA algorithm. The method as described in item 46 of the patent application scope, wherein the aforementioned output data corresponds to multiple files. The method as described in item 50 of the patent application scope further includes extracting the plurality of files from the output data through the TAR algorithm. The method as described in any of items 36-51 of the patent application range, wherein the aforementioned 5-bit transcoding framework is based on Table 2. The method as described in any one of patent application items 36-53, wherein the nucleic acid set includes primer sequences at the 3'and 5'ends and the method includes removing the primer sequence before the nucleotide decoding step . A method for retrieving output data stored on a nucleic acid group of interest, characterized in that the aforementioned nucleic acid group of interest is one of multiple sets of nucleotide sequences present in the mixture, each group encoding Different output data sets with different primer pair sets at the 3'and 5'ends, the aforementioned method includes: a) amplifying the aforementioned nucleic acid set using primer pairs corresponding to the nucleic acid of interest; b) obtaining the amplified nucleic acid Nucleotide sequence group, c) converting the aforementioned nucleotide sequence group into the aforementioned output data according to the method described in any of items 40-52 of the patent application scope; thereby obtaining the aforementioned output data. A method for retrieving corresponding two or more sets of output data stored on two or more sets of nucleic acids of interest, characterized by the fact that the aforementioned two or more sets of nucleic acids of interest exist Among the multiple nucleotide sequences in the mixture, each set encodes a different set of output data and has different sets of primer pairs at the 3'and 5'ends. The foregoing methods include: a) using the corresponding two sets Or more pairs of primers of the nucleic acid of interest to amplify the aforementioned two or more sets of nucleic acids of interest; b) obtain two or more sets of nucleotide sequences of the amplified nucleic acid, c) if the patent application The method described in any one of items 40-52 converts the aforementioned two or more sets of nucleotide sequences into the aforementioned two or more sets of output data; thereby obtaining the aforementioned two or more sets of output data. A non-transitory computer-readable storage medium storing one or more programs, characterized in that the aforementioned one or more programs include instructions that when executed by one or more processors of an electronic device, cause the aforementioned The electronic device implements the method described in any one of the patent application items 2-36 or 40-52. A system for providing nucleic acid-based data storage or retrieving data from nucleic acid, characterized in that it includes: one or more processors; memory; and one or more programs, wherein the aforementioned one or more programs Stored in the aforementioned memory and configured to be executed by the aforementioned one or more processors, the aforementioned one or more programs including are used to implement as described in any one of patent application scope 2-36 or 40-52 Method instructions. An electronic device for providing nucleic acid-based data storage or retrieving data from a nucleic acid, characterized in that the aforementioned device includes a device for implementing as described in any one of patent application items 2-36 or 40-52 Method equipment.

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