WO2024027620A1 - Data storage medium and use thereof - Google Patents

Abstract

Provided in the present application are a data storage medium and a use thereof. Also provided is a nucleic acid molecule. The nucleic acid molecule can be combined with a carrier having addressable information, and data information contained in the nucleic acid molecule can be randomly read, erased or written in situ in the carrier.

Description

A data storage medium and its application Technical field

This application relates to the field of data storage, and specifically to a data storage medium and its application.

Background technique

With the rise of big data and artificial intelligence technology, the demand for storage of massive data has also shown explosive growth, and mainstream storage media are gradually unable to meet the rapidly growing storage demand. Deoxyribonucleic acid (DNA), as the storage medium for the carbon-based genetic code of life selected by hundreds of millions of years of natural evolution, has extremely high storage density and robustness. The codability and efficient replication capabilities of DNA may provide a new strategy for high-density data storage and high-performance computing. DNA storage has the advantage of high physical stability, unlike electronic media that deteriorates with the number of reads, providing a fundamental solution for long-term data storage. In addition, DNA has both information processing and computing capabilities, providing new ideas for the development of new storage-computing integrated architectures and systems.

The DNA data storage process mainly includes six technical links: information encoding, writing, saving, retrieval, reading, and decoding. The existing DNA storage system is mainly based on artificially synthesized short-chain DNA or long-chain DNA, which encodes digital information into DNA sequences and synthesizes corresponding DNA strands, which are stored in cells or outside the body, and then sequenced technology is used to read out the data.

Compared with existing mature data storage systems, existing DNA storage systems mainly include two storage modes. One storage mode is a non-random read storage architecture. This solution encodes and writes the data to be stored holistically. Therefore, the reading of the data also requires sequencing of all sequences in the system and the overall sequencing results. Decode to obtain data information. The non-random read storage architecture cannot search for partial information within the file, and therefore cannot perform addressable modifications to the written information. Another storage mode is a DNA storage architecture with a random read mode. In this type of system, the data to be written is first divided into data fragments, the data fragments are encoded and index sequences are added. Therefore, this type of DNA storage architecture has the ability to selectively read information fragments. However, the selective reading of data in this type of system mainly relies on PCR technology. It is necessary to directly take out a part of the original data or extract the data to be read through magnetic beads and then perform PCR amplification, and then perform sequencing reading. However, this method will destroy the original data composition. After multiple reads, multiple PCRs will introduce many errors into the data, affecting the recovery of the original data. Similar to the in-situ reading method of traditional semiconductor storage media, a data reading method that achieves selective reading of data without destroying the original data is still lacking. In addition, the current DNA storage system mainly has a single write archiving function, but the ability to modify data in the storage system after writing is still very insufficient, especially the addressable modification of any data stored in the DNA storage system. Still not implemented.

Therefore, there is an urgent need in the art for a DNA storage method that is addressable to write, addressable to modify, and/or addressable to read.

Contents of the invention

This application provides a DNA storage system with complete data operation capabilities, which can realize data writing, data deletion, data modification, data reading and other addressable features, especially multiple erasing, data modification and multiple Secondary data readout can make up for the functional shortcomings of existing DNA storage systems. For example, this application enables programmable combination and separation of storage addresses and data on DNA molecules.

In one aspect, the present application provides a system comprising 1) a nucleic acid molecule containing data information, and 2) a carrier having addressable information, the nucleic acid molecule being able to bind to the carrier, and the The data information contained in the nucleic acid molecules can be stored (also called writing), read in situ and/or edited in situ (including erasing and/or writing new data information) on the carrier.

On the other hand, the present application provides a method for data storage, which method includes providing 1) the nucleic acid molecule containing data information, and 2) the carrier with addressable information.

On the other hand, the present application provides a method for data reading, which method includes providing 1) the nucleic acid molecule containing data information, and 2) the carrier with addressable information.

On the other hand, the present application provides a method for data editing, which method includes providing 1) the nucleic acid molecule containing data information, and 2) the carrier with addressable information, and replacing the nucleic acid molecule data information in.

In certain embodiments, the carrier has different sequences of address sequences at different physical locations.

In certain embodiments, the vector comprises a DNA origami substrate comprising a staple strand comprising an address sequence having addressable information.

In some embodiments, in the vector, the address sequences at the ends of the staple chains are arranged in a matrix. The matrix is at least a 2×2 matrix. In some embodiments, the distance between two adjacent staple strands is about 6-24 nm. For example, the distance between two adjacent staple chains is approximately 6 nm. For example, the distance between two adjacent staple strands is approximately 12 nm. For example, the distance between two adjacent staple strands is approximately 18 nm. For example, the distance between two adjacent staple strands is approximately 24 nm.

In certain embodiments, the nucleic acid molecules are bound to the staple strands and arranged in a matrix. The matrix is at least a 2×2 matrix. In certain embodiments, the distance between two adjacent nucleic acid molecules is about 6-24 nm. For example, the distance between two adjacent nucleic acid molecules is approximately 6 nm. For example, the distance between two adjacent nucleic acid molecules is approximately 12 nm. For example, the distance between two adjacent nucleic acid molecules is approximately 18 nm. For example, the distance between two adjacent nucleic acid molecules is approximately 24 nm.

In certain embodiments, the nucleic acid molecule comprises an address complementary sequence that is complementary to the address sequence on the vector. The nucleic acid molecule is bound to a specific physical location of the vector by complementarity between the address complementary sequence and the address sequence.

In certain embodiments, the address complement is about 15 or more nucleotides in length.

In some embodiments, the nucleic acid molecule includes a data sequence, and the data information in the data sequence can be read in a state where the nucleic acid molecule and the vector are not substantially separated. For example, by activating a reaction, a readable data chain is synthesized, thereby reading the data information in the data sequence.

In certain embodiments, the data sequence is about 1 or more nucleotides in length. The data sequence may be single stranded.

In some embodiments, the nucleic acid molecule includes a read priming sequence, the read priming sequence can trigger the synthesis or transcription of the data sequence into a strand to be sequenced, and the strand to be sequenced is complementary to the data sequence. .

In certain embodiments, the read priming sequence includes a promoter. For example, the read initiating sequence is a T7 promoter. The read priming sequence enables transcription of the data sequence. By sequencing and decoding the transcript products, the data information contained in the data sequence can be read.

In some embodiments, the system and/or method further includes 3) a blocking chain that can prevent the data information from being read. In this application, the blocking chain may also be called a closed chain.

In some embodiments, the blocking strand includes a blocking sequence that is complementary to the read initiating sequence, thereby preventing the data information from being read. In a specific embodiment, the blocking sequence includes a promoter complementary sequence that binds to the promoter as a read initiator sequence, so that transcription cannot be initiated.

In certain embodiments, the blocking strand includes a blocking elongation sequence located upstream and/or downstream of the blocking sequence, and the blocking elongation sequence is substantially non-complementary to the nucleic acid molecule. In this application, the elongation-blocking sequence may also be referred to as a dangling chain (teohold). Dangling strands can provide energy driving force and specificity for transcriptional activation reactions. In a specific embodiment, the elongation-blocking sequence is about 4-10 nucleotides in length, for example, 6 nucleotides.

In this application, for the data information at each address, in the storage state or when no reading is required, the blocking chain is complementary to the read initiation sequence on the nucleic acid molecule, and transcription cannot be initiated.

In some embodiments, the system and/or method further includes 4) activation chain. For example, when data reading and writing are required, the activation chain can be added. The activation chain can prevent the blocking chain from binding to the nucleic acid molecule, and the activation chain is complementary to the blocking sequence and the blocking extension sequence at the same time. The activation chain corresponds to a specific physical address. When the data information of a specific address needs to be read, an activation chain corresponding to the specific address can be added. The activation chain combines with the hindrance chain, causing the hindrance chain to detach from the reading initiation sequence of the nucleic acid molecule. , thereby initiating reading, for example, activating the transcription function at that address. In a specific embodiment, an activation chain corresponding to a specific address is added to activate the transcription function of the address. Under the action of T7 RNA polymerase, the data of the address is transcribed into an RNA chain. The unactivated address has no transcription function. ability. By collecting the obtained RNA strands for subsequent sequencing, the nucleic acid sequence is decoded into data information, thereby achieving addressable data. Read.

In some embodiments, the data reading process can be performed at room temperature. At room temperature, the blocking strand bound to the nucleic acid molecule at a specific physical location can bind to the activating strand.

In certain embodiments, the nucleic acid molecule includes an erasure function sequence, the erasure function sequence is located upstream and/or downstream of the address complementary sequence, and the erasure function sequence is consistent with all the addresses on the carrier. The above address sequences are basically not complementary.

In some embodiments, the system and/or method further includes 5) erasure chain. For example, when data erasure is required, the erasure chain can be added. The erasure strand can prevent the nucleic acid molecule from binding to the carrier, and the erasure strand can be complementary to the address complementary sequence and the erasure function sequence at the same time. In certain embodiments, the erasing strand is capable of complementary binding to the erasing functional sequence of the nucleic acid molecule. The erasure chain corresponds to a specific physical address. When the data information of a specific address needs to be erased, the erasure chain corresponding to the specific address can be added, and then, for example, through a DNA chain substitution reaction, the data sequence of the address is combined, so that the data sequence Removed from the carrier, returning the address to its unwritten state.

In some embodiments, the erasing process can be performed at room temperature. At room temperature, after the erasure strand is added, the nucleic acid molecules at specific physical locations can bind to the erasure strand, and the nucleic acid molecules are basically not bound to the carrier.

In some embodiments, when the original data information is erased, that is, when the original data sequence is erased by the erasure chain, the address can be rewritten into a new data sequence, for example, adding new data information containing nucleic acid molecules. This enables in-situ modification and/or editing of data.

In certain embodiments, the molar ratio of the nucleic acid molecule to the address sequence in the vector is about 1:1 or greater. In certain embodiments, the molar ratio of the nucleic acid molecule to the address sequence in the vector is about 2:1 or greater. In certain embodiments, the molar ratio of the nucleic acid molecule to the address sequence in the vector is about 3:1 or greater. In certain embodiments, the molar ratio of the nucleic acid molecule to the address sequence in the vector is about 4:1 or greater. In certain embodiments, the molar ratio of the nucleic acid molecule to the address sequence in the vector is about 5:1 or greater.

In another aspect, the application provides a nucleic acid molecule in the system and/or method, the nucleic acid molecule comprising the data sequence, address complement sequence and/or read priming sequence. In one embodiment, upstream and/or downstream of the data sequence, the nucleic acid molecule further includes primers for verifying the feasibility of erasing and reading. In other specific embodiments, the primer part can be replaced with a data sequence containing data information to further increase storage capacity. Figure 2 shows an example of a nucleic acid molecule described herein.

In another aspect, the application provides vectors in the systems and/or methods.

On the other hand, the present application provides a storage medium, which contains the method of the present application.

On the other hand, the present application provides a device, the device includes the storage medium of the present application, and is coupled to the A processor of a storage medium, the processor is configured to execute based on a program stored in the storage medium to implement the method of the present application.

In a specific embodiment, the inventor of the present application constructed a 6nm DNA chip to achieve multiple in-situ erasing and/or multiple in-situ selective reading (WMRM). In a more specific implementation, the addressability and programmability of DNA nanostructures are used to arrange data sequences storing information on DNA origami with a pitch of 6 nm, giving each data sequence a physical structure with a pitch of 6 nm. Address, by designing erasure chains and utilizing the information processing and computing capabilities provided by the DNA strand displacement reaction, can realize repeated erasing of DNA data sequences on origami and modify any data. In addition, a promoter (for example, T7 promoter) switch is set on the DNA chain as a read start sequence. By designing a blocking chain and/or an activating chain, the DNA chain displacement reaction is used to regulate the transcriptional activity of the DNA chain, and the transcription activity of the DNA chain is selectively The DNA information is transcribed and RNA sequenced (for example, using Illumina technology) to achieve selective reading and multiple reading of the data information.

Compared with the existing technology, this application at least has the following characteristics: it is based on DNA assembly technology and has nanoscale addressability, which is not available in the current DNA storage system; using this nano-addressability, it can The area density of DNA storage has been increased to 1 bit/square nanometer, surpassing existing inorganic storage architecture and DNA storage architecture; based on the controllable dynamic assembly of the DNA origami surface, a full-featured storage system is realized, including data reading, writing, modification, etc. Operation; the readout process adopts a solution of in-situ transcription into RNA molecules and sequencing of the transcript products without changing the original storage system, achieving non-destructive DNA data readout. For example, the method and data carrier of the present application can have the effect of multiple reading and writing, and multiple reading. For example, after reading the data system of the present application multiple times, the structure of the data system basically does not change. For example, the address information of the data carrier of the present application has high accuracy and specificity, and can achieve a higher level of discrimination. For example, the method and data system of the present application can realize the reading of data information at room temperature to about 42 degrees Celsius.

Those skilled in the art will readily appreciate other aspects and advantages of the present application from the detailed description below. Only exemplary embodiments of the present application are shown and described in the following detailed description. As those skilled in the art will realize, the contents of this application enable those skilled in the art to make changes to the specific embodiments disclosed without departing from the spirit and scope of the invention covered by this application. Accordingly, the drawings and descriptions of the present application are illustrative only and not restrictive.

Description of the drawings

The specific features of the invention to which this application relates are set forth in the appended claims. The features and advantages of the invention to which this application relates can be better understood by reference to the exemplary embodiments described in detail below and the accompanying drawings. A brief description of the drawings is as follows:

Figure 1 shows an exemplary operation flow of the data storage medium of the present application. For example, the stored data can be used for all data forms, including but not limited to Chinese characters; the storage capacity can be infinitely expanded based on the length of the data sequence, including but not limited to 16 bits/site or 120bits/site; for data reading, sequencing technologies known in the art can be used, including but not limited to high-throughput in-situ sequencing, transcription-RNA sequencing, and transcription-reverse transcription-amplification- DNA sequencing.

Figure 2 shows an exemplary structural composition of the nucleic acid molecule of the present application, that is, the data chain includes the data sequence, in which primer 1 and primer 2 are designed only to more easily verify the feasibility of selective erasing and reading, This part can be replaced with actual stored information to further increase storage capacity.

Figure 3 shows an exemplary process of selective reading and reversible erasing of the data storage medium of the present application. Transcription activation process: the key chain is the activation chain; when the nucleic acid molecule described in this application is blocked by the hindrance chain, transcription cannot be started. When the key chain is added, the hindrance chain turns to combine with the key chain and no longer binds to the promoter, adding After the T7 promoter is intact, RNA polymerase binds to the promoter and initiates transcription. Reversible erasing: When the erasing chain (ie erasure chain) is added, the data chain is combined with the erasing chain and separated from the carrier. After rejoining the complementary sequence data chain with the same address, the data chain is re-written to the carrier.

Figure 4 shows the structural characterization results after writing DNA origami surface data. The upper part is a schematic diagram of the address where data is written on the surface of DNA origami. The lower part shows the AFM characterization results after data writing.

Figure 5 shows a heat map of PAGE data statistics for readout activation orthogonality. Only when the activation chain matches the data chain can the data be read out efficiently.

Figure 6 shows the second-generation sequencing results after seven address data reads. The sequence information obtained by sequencing at each address is completely consistent with the written information.

Figure 7 shows the super-resolution fluorescence microscopy imaging results of addressed data erasing. The upper part shows the data fixed-point erasure and writing process, as well as the corresponding data address schematic diagram. The figure below shows the fluorescence imaging structural characterization results for each step of the process.

Figure 8 shows the fluorescence test results of the feasibility of repeated data modification.

Figure 9 shows a single-point TIRF (total internal reflection fluorescence microscope) imaging image of seven address data being repeatedly erased 10 times.

Figure 10 shows the TIRF (total internal reflection fluorescence microscopy) imaging and co-localization statistical chart of seven address data that were repeatedly erased 10 times. The co-localization of red and green fluorescence represents the presence of data on the origami paper.

Figure 11 shows the qPCR quantification of transcribed RNA during repeated reads of the data chain.

Figure 12 shows the writing results of the data link under four different matrix spacings.

Figure 13 shows the readout results of the data link under four different matrix spacings.

Detailed ways

The implementation of the invention of the present application will be described below with specific examples. Those familiar with this technology can easily understand other advantages and effects of the invention of the present application from the content disclosed in this specification.

Definition of Terms

In this application, the term "addressability" generally refers to associating specific information with a location on a storage medium. For example, in order to selectively access, read, and/or modify information at a specific location, addressability of the data carrier is required. For example, when recording data information, the information carrier can simultaneously record basically uniquely corresponding index information (address) corresponding to different data information. For example, the address information of specific data information is recorded through the physical location, spatial location, etc. of the data carrier. For example, when the data carrier is addressable, information desired to be accessed, read, and/or modified may be selectively accessed and may not need to be accessed one by one.

In this application, the terms "nucleic acid molecule," "nucleic acid sequence," and "nucleic acid fragment" are used interchangeably and generally refer to deoxyribonucleotides or ribonucleotides of various lengths, or analogs thereof. Exemplary nucleotides include deoxyribonucleotides (DNA) or ribonucleotides (RNA), or non-standard nucleotides, nucleotide analogs and/or modified nucleotides.

In this application, the term "vector" generally refers to a substance capable of carrying nucleic acid molecules. For example, the vector may comprise nucleic acid nanostructures, which may be made from nucleic acids such as DNA, RNA, locked nucleic acid (LNA), peptide nucleic acid (PNA), or any combination thereof. Two- or three-dimensional nanostructures. For example, single-stranded nucleic acids or double-stranded nucleic acids (eg, having only a helical structure) may not be considered "nanostructures." In some embodiments, nucleic acid nanostructures serve as scaffolds for the formation of more complex structures, such as molecular complexes. In some embodiments, the nucleic acid nanostructure is a DNA origami structure assembled using DNA origami methods. For example, nucleic acid origami nanostructures may refer to nucleic acid nanostructures formed by assembling two or more "staple strands" with one or more "scaffold" strands into a prescribed shape. Staple strands are typically short (eg, 50 nucleotides or less) nucleic acid strands (single-stranded nucleic acids); scaffold strands are typically longer (eg, longer than 200 nucleotides) nucleic acid strands (single-stranded nucleic acids). nucleic acids). The nucleic acid origami nanostructure may be a DNA origami nanostructure.

DNA origami nanostructures can fold (e.g., through self-assembly) into discrete and unique geometric patterns, such as two-dimensional (2D) and three-dimensional (3D) shapes, which can further self-assemble to create structures containing two or more discrete origami nanostructures. Larger nanostructures or microstructures of structures. In some embodiments, the scaffold strand has sequences derived from M13 phage. Other bracket chains can be used. In some embodiments, the staple strands are fluorophore labeled staple strands. In some embodiments, the staple strand is 4 to 30 nucleotides in length (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 nucleotides). In some embodiments, the staple strands are stably bonded to the scaffold strands (for longer than 10 seconds), for example, at room temperature. In some embodiments, the staple strands are stably bonded to the scaffold strands (for longer than one week), for example, at room temperature. In some embodiments, the staple strand is greater than 30 nucleotides in length.

In this application, the term "in situ" generally refers to operations performed in the original location. For example, in-situ reading refers to The nucleic acid molecules that record the data are read at the original position of the carrier, without the need to release the nucleic acid molecules into the solution first and then read the data. The term "amplification" is generally intended to include the production of copies of a nucleic acid molecule via repeated cycles of primed enzymatic synthesis. The reading step of the present application may include a polymerization step, including but not limited to polymerase chain reaction (PCR), transcription to RNA, and the like, including, for example, any other nucleic acid amplification and/or transcription known to those skilled in the art. Technology.

As used herein, the term "complementary" or "complementarity" generally refers to the ability of a nucleic acid to form hydrogen bonds with another nucleic acid sequence via traditional Watson-Crick or other non-traditional types. For example, the sequence A-G-T is complementary to the sequence T-C-A. Percent complementarity indicates the percentage of residues in a nucleic acid molecule that can form hydrogen bonds (e.g., Watson-Crick base pairing) with a second nucleic acid sequence (e.g., 50 out of 10, 6, 7, 8, 9, 10, respectively) %, 60%, 70%, 80%, 90% and 100% complementary). For example, "perfectly complementary" means that all contiguous residues of a nucleic acid sequence will hydrogen bond to the same number of contiguous residues in a second nucleic acid sequence. For example, "substantially complementary" means that at 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, Within a region of 40, 45, 50 or more nucleotides, or refers to at least 60%, 65%, 70%, 75%, 80%, A degree of complementarity of 85%, 90%, 95%, 97%, 98%, 99% or 100%. For example, "binding" generally refers to a species that uniquely binds to a particular species, such as in a sequence-specific manner.

In this application, the term "comprising" generally means the inclusion of explicitly specified features, but not the exclusion of other elements.

In this application, the term "about" generally refers to a variation within the range of 0.5% to 10% above or below the specified value, such as 0.5%, 1%, 1.5%, 2%, 2.5%, above or below the specified value. 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%.

Detailed description of the invention

On the one hand, the present application provides a nucleic acid molecule that can be bound to a carrier with addressable information, and the data information that the nucleic acid molecule can contain can be randomly read and erased in situ on the carrier. . For example, the nucleic acid molecules provided in this application can be used as a data link. The data link can be combined with a specific position on the carrier based on the principle of base complementarity. Combined with dynamic DNA assembly technology, programmable combination and separation of storage addresses and data can be achieved. For example, when the data information is read, the original storage system does not need to be changed, achieving non-destructive data reading. For example, when reading the data information recorded in the nucleic acid molecule, the nucleic acid molecule does not need to be released from the carrier first, thereby achieving in-situ random reading of the required read information.

For example, address sequences at different physical locations on the carrier have different sequences, and the nucleic acid molecule may include an address complementary sequence, and the address complementary sequence is complementary to the address sequence on the carrier. For example, the address complementary sequence can be an address recognition sequence, and the address complementary sequence can recognize and specifically bind the address sequence on the carrier. For example, the address information can be recorded by a sequence of addresses at different physical locations on the carrier. For example, there can be 2 or more coordinate points on the carrier. For example, different coordinate points represent different physical locations. For example, specific coordinate points can be located by the ends of staple strands at specific locations in DNA origami. For example, different coordinate points extend an address sequence with a unique sequence composition. For example, the address sequence of this application may be a linear single chain or a branched chain structure. For example, the address sequence of a specific physical location can be used to record index information because it has a unique sequence. When a substantially unique complementary address complementary sequence (contained by a specific data link) is combined with the address sequence, The data information may be combined with the address information. In some embodiments, the data link is stably associated with the address link (eg, for longer than 10 seconds), such as at room temperature. In some embodiments, such as at low temperatures (about 4 degrees Celsius), the data link is stably coupled to the address chain (eg, for longer than 10 seconds). In some embodiments, for example, stored in a dry powder state on a bacteriostatic, antioxidant, and high temperature resistant plate, the data link and the address link are stably combined (eg, longer than 10 seconds).

For example, the address complementary sequence is about 15 or more nucleotides in length. For example, the address complementary sequence of the present application may be a linear single chain or a branched chain structure. For example, the length of the address complementary sequence may be about 10 or more, about 11 or more, about 12 or more, about 13 or more, about 14 or more, about 15 or more, about 16 or more, about 17 or more, about 18 or more, about 19 or more, about 20 or more, about 25 or more, about 30 or more, about 40 or more, about 50 or more, or about 100 or more nucleotides. For example, the nucleotides may include natural nucleotides and/or nucleotides with artificial modifications, such as, but not limited to, methyl modifications, amino modifications, fluoro modifications, and the like.

For example, the nucleic acid molecule may further comprise an erasure function sequence, the erasure function sequence is located upstream and/or downstream of the address complementary sequence, and the erasure function sequence is identical to the address sequence on the carrier. Basically not complementary. For example, the erase and write function sequence may also be called an erase and write functional area. For example, the erase and write function sequence is located upstream and/or downstream of the address complementary sequence. For example, the sequence composed of the erasing function sequence and the address complementary sequence may not be completely complementary to the address sequence on the carrier. For example, the exemplary address complementary sequence may be 20 nucleotides, and the erase function sequence may be 10 nucleotides. The address sequence on the carrier may only be complementary to the 20 nucleotides of the address complementary sequence, but may not be complementary. The erasing function sequence and the address complementary sequence together form 30 nucleotides that are completely complementary. For example, a strand with a higher binding capacity (such as the erasure strand of the present application) can thus be introduced so that the nucleic acid sequence of the data strand can be removed (erased) from a specific position of the vector.

For example, when the erasure chain exists, the nucleic acid molecule can not be combined with the carrier, and the erasure chain is complementary to the address chain and the erasure function sequence at the same time. For example, at room temperature, the nucleic acid molecule (data strand) of the present application can have stronger binding ability to the erasure strand. For example, the data strand may have more and/or stronger binding bases to the erasure strand than to the address sequence on the carrier.

For example, in order to achieve addressable erasing and writing, the address complementary sequence of the nucleic acid molecule (data chain), the address sequence corresponding to a specific position of the carrier, and the sequence of the specific corresponding erasure chain can be specified through design. A sequence of base sequences that has a unique complementary matching pattern. For example, the address sequence of a specific position of the vector is uniquely complementary to the address complementary sequence of a specific nucleic acid molecule (data link) to achieve addressable writing. For example, the address complementary sequence of a specific nucleic acid molecule (data strand) bound to a specific position of the carrier is uniquely complementary to the sequence of the specific corresponding erasure strand to achieve addressable erasure.

For example, the nucleic acid molecule may include a data sequence, and the data information in the data sequence may be read in a state where the nucleic acid molecule and the carrier are not substantially separated. For example, when the nucleic acid molecules (data links) are in the stored position, the data information can be read. For example, reading methods include but are not limited to in-situ sequencing, in-situ transcription into other nucleic acid molecules for reading information, in-situ reading into other nucleic acid molecules for reading information, and so on. For example, in this application, deriving information stored in a nucleic acid molecule (data link) can be considered as reading. For example, sequencing the derived (eg transcribed/amplified) other nucleic acid molecules may be considered a subsequent optional additional read step.

For example, the data sequence is about 1 or more nucleotides in length. For example, the data sequence of this application may be a linear single chain or a branched chain structure. For example, the length of the data sequence may be about 1 or more, about 2 or more, about 3 or more, about 4 or more, about 5 or more, about 6 or more, about 7 or more, about 8 or more, about 9 or more, about 10 or more, about 15 or more, about 20 or More, about 30 or more, about 40 or more, about 50 or more, about 100 or more, about 120 or more, about 150 or more , about 200 or more, about 500 or more, about 700 or more, or about 1000 or more. For example, the storage method of this application can be compatible with data sequences of any length.

For example, the nucleic acid molecule may include a read priming sequence capable of priming the data sequence to be read as a strand to be sequenced, and the strand to be sequenced is complementary to the data sequence. For example, the nucleic acid molecule may comprise a read priming sequence that may cause the data sequence to be read. For example, the read priming sequence of the present application can be a promoter, such as including but not limited to T7 promoter. For example, the storage method of the present application can be compatible with read priming sequences of any length and type, and the read priming sequences have specific sequences that can be used for binding and polymerization initiation of polymerases known in the art.

For example, the nucleic acid molecule (data link) of the present application can have the function of selective reading. For example, the selective reading can be achieved by introducing a blocking strand whose blocking sequence is complementary to the reading initiating sequence. For example, when a blocking chain exists, the data sequence cannot be read, the blocking chain may include a blocking sequence, and the blocking sequence of the blocking chain is partially or completely complementary to the read initiating sequence. For example, the blocking sequence of the blocking strand is complementary to the read priming sequence and an interval of about 5 nucleotides upstream/downstream of the reading priming sequence. For example, the blocking sequence is long The length is 22 nucleotides, of which 17 nucleotides are complementary to the read priming sequence, and the other 5 nucleotides are complementary to an interval of approximately 5 nucleotides upstream/downstream of the read priming sequence. For example, the blocking chain has a higher ability to bind to the data chain than the T7 promoter binds to the data chain. For example, the blocking chain of the present application can be combined with the read trigger sequence of the nucleic acid molecule (data chain), the data chain, or any position that can block the derivation of the information stored in the nucleic acid molecule (data chain).

For example, the blocking strand may further comprise a blocking extension sequence located upstream and/or downstream of the blocking sequence, and the blocking extension sequence is substantially not complementary to the nucleic acid molecule. For example, the blocking chain has a blocking sequence and a blocking extension sequence. When the blocking chain is combined with the nucleic acid molecule (data chain), the blocking extension sequence is basically not combined with the nucleic acid molecule (data chain), for example, forming an overhang. structure. For example, the extension-blocking sequence may be about 8 nucleotides in length. For example, the blocking extension sequence can be used as a lever by improving the binding ability of the activation chain and the blocking chain, so that the activation chain removes the blocking chain from the data chain.

For example, the blocking strand can not bind to the nucleic acid molecule when an activating strand is present, which is complementary to the blocking sequence and the blocking extension sequence at the same time. For example, during the annealing process, the activation chain of the present application can have stronger binding ability with the hindrance chain. For example, the activating strand may have more and/or stronger binding bases to the blocking strand than to a sequence on the nucleic acid molecule (data strand) that binds to the blocking strand.

For example, in order to achieve addressable reading, the sequence of the nucleic acid molecule (data chain) used to block the reading, the blocking sequence of the specific corresponding blocking strand, and the sequence of the specific corresponding activating chain can be designed by designing specific bases. A sequence of base sequences with a unique complementary matching pattern. For example, the sequence of the nucleic acid molecule (data strand) used to block the read is uniquely complementary to the blocking sequence of the specific corresponding blocking strand to achieve an addressable locked read. For example, the sequence of a particular corresponding activating strand is uniquely complementary to the sequence of the blocking sequence of a particular corresponding blocking strand to enable addressable unlocking (activating) reading. For example, when a specific nucleic acid molecule (data link) is in a locked reading state, the data sequence of the specific nucleic acid molecule (data link) may not be substantially read, transcribed and/or amplified. For example, when a specific nucleic acid molecule (data link) is in an unlocked (activated) reading state, the data sequence of the specific nucleic acid molecule (data link) can be read, transcribed and/or amplified.

For example, the vector may comprise a DNA origami substrate whose staple strands may contain address sequences with addressable information. For example, there can be 2 or more coordinate points on the carrier. For example, different coordinate points represent different physical locations. For example, specific coordinate points can be located by the ends of staple strands at specific locations in DNA origami. For example, different coordinate points extend an address sequence with a unique sequence composition. For example, the address sequence of a specific physical location can be used to record index information because it has a unique sequence. When a substantially unique complementary address complementary sequence (contained by a specific data link) is combined with the address sequence, The data information may be combined with the address information. For example, 2 or more of the staple strands are spaced apart by about 6 nanometers or more. For example, the adjacent staples The strands are spaced about 6 nanometers or greater, about 7 nanometers or greater, about 8 nanometers or greater, about 9 nanometers or greater, about 10 nanometers or greater, about 15 nanometers or greater, about 20 nanometers or more. larger, about 25 nanometers or larger, or about 30 nanometers or larger.

The present application provides a system, which may include the nucleic acid molecule of the present application, and a vector. For example, the system may also include the erasure chain of the present application, the blocking chain of the present application, and/or the activation chain of the present application.

The present application provides a data storage method, which may include providing the nucleic acid molecule of the present application and/or the system of the present application. A data editing and/or data reading method, the data editing method may include replacing the data information stored in the nucleic acid molecules of the present application. A data reading method, which may include determining the data information stored in the nucleic acid molecules of the present application.

For example, the method may further comprise providing a vector, the vector may comprise a DNA origami substrate, the staple strands of the DNA origami may comprise an address sequence having addressable information, and the method may comprise providing a molar ratio of About 2:1 or higher of the nucleic acid molecule to the address sequence. For example, the method may include providing a molar ratio of about 2:1 or higher, about 2.1:1 or higher, about 2.2:1 or higher, about 2.3:1 or higher, about 2.4:1 or higher. , about 2.5:1 or higher, about 3:1 or higher, about 4:1 or higher, about 5:1 or higher, about 10:1 or higher, about 20:1 or higher, about 50:1 or higher, or about 100:1 or higher of the nucleic acid molecule to the address sequence. For example, choosing an appropriate molar ratio can improve the storage success rate of the data link. For example, choosing an appropriate molar ratio can improve the storage cost-effectiveness of the data link.

For example, the method may further comprise providing an erasure strand of the present application, the nucleic acid molecule at a specific physical location is bound to the erasure strand at room temperature, and the nucleic acid molecule is not substantially bound to the carrier. The terms "room temperature" and "ambient temperature" generally refer to a temperature between about 16 degrees Celsius and about 40 degrees Celsius. For example, a temperature between about 16 degrees Celsius and about 25 degrees Celsius. For example, about 25 degrees Celsius.

For example, the method may further include providing a blocking chain of the present application, the blocking chain is combined with the nucleic acid molecule, and the data information of the nucleic acid molecule cannot be substantially read. For example, the method may further include providing the activation chain of the present application. During the heating and cooling processes, the hindrance chain to which the nucleic acid molecules at a specific physical position are bound binds to the activation chain, and the hindrance chain does not substantially bind to the activation chain. The nucleic acid molecules bind. For example, the heating and cooling processes in this application include the process of annealing nucleic acids. For example, the process of partially separating the double-stranded nucleic acid molecules of the present application (eg, heating) and then restoring the partially double-stranded structure (eg, cooling) can be a nucleic acid annealing process. For example, the process of heating to about 95 degrees for about 3 minutes and then cooling to room temperature at a rate of about 1.2 degrees per minute can be a nucleic acid annealing process.

On the other hand, the present application provides a storage medium recording a program that can run the method of the present application.

On the other hand, the present application provides a device, which may contain the storage medium of the present application. On the other hand, the present application provides a non-volatile computer-readable storage medium on which a computer program is stored, and the program is executed by a processor to implement any one or more methods described in the present application. For example, the non-volatile computer-readable storage medium may include software disk, flexible disk, hard disk, solid state storage (SSS) (such as solid state drive (SSD)), solid state card (SSC), solid state module (SSM)), enterprise flash drive, tape or any other non-transitory magnetic media, etc. Non-volatile computer-readable storage media may also include punched cards, paper tape, cursor pads (or any other physical media having a hole pattern or other optically identifiable markings), compact disk read-only memory (CD-ROM) , Compact Disc Rewritable (CD-RW), Digital Versatile Disc (DVD), Blu-ray Disc (BD) and/or any other non-transitory optical media.

For example, the device of the present application may further include a processor coupled to the storage medium, and the processor is configured to execute based on a program stored in the storage medium to implement the method of the present application. For example, the device may implement various mechanisms to ensure that methods described herein when executed on a database system produce correct results. In this application, the device may use disks as permanent data storage. In this application, the device can provide database storage and processing services for multiple database clients. The device may store database data across multiple shared storage devices and/or may utilize one or more execution platforms with multiple execution nodes. The device can be organized so that storage and computing resources can be expanded effectively infinitely.

This application provides the following implementation methods:

The preparation of a nanometer-addressable, fully functional DNA storage system includes the following steps:

(1) Design and assemble nano-addressable DNA platform. The address on the DNA origami surface is expressed in the form of a DNA sequence. An orthogonal sequence library with a length of, for example, 15 to 18 bases or longer is designed through a sequence screening algorithm to ensure that the data chain can be stored in a stable hybridization manner. to a specific address. Using a single-layer rectangular DNA origami as a template, the address sequence is extended from the corresponding site on the origami surface by designing its staple strands. The address-designed staple strand combination is mixed with the backbone strand and assembled through annealing to form an addressable blank DNA storage platform.

The DNA origami assembly process can refer to the DNA origami assembly technology known in the art, such as Rothemund, P.W.K. Folding DNA to Create Nanoscale Shapes and Patterns. Nature 2006, 440 (7082), 297–302.

An exemplary overall storage process can be shown in Figure 1, an exemplary data chain (nucleic acid molecule in this invention) structure can be shown in Figure 2, and selective reading and reversible erasing of data can be shown in Figure 3.

(2) Encode the data corresponding to the address. Divide the data to be written into fragments that match the capacity of a single address, add the T7 promoter sequence and address sequence to form a data chain sequence to be written, and synthesize the data chain. Methods of synthesizing nucleic acid strands are well known in the art, for example, by chemically synthesizing data strands.

A data operation system of an addressable DNA storage system, the system may include:

(1) Data writing (or storing) step, which may include utilizing the DNA complementary pairing principle to add a data chain containing a data sequence to bind it at a specific address.

(2) Data erasure step, which may include adding a specific address erasure operation DNA chain (also known as Erase chain, that is, the erasure chain), through the DNA chain substitution reaction, combined with the data sequence there, the address is restored to an unwritten state.

(3) Data modification (or editing) step, which may include first erasing the original data and then writing new data.

(4) Data readout (or reading) step, which may include adding an activation chain (also called an Input chain) corresponding to a specific address to activate the transcription function of the address, under the action of T7 RNA polymerase , transcribe the data of this address into an RNA chain, and the inactive address does not have the transcription ability. Addressable data reading is achieved by collecting the obtained RNA strands for subsequent sequencing.

Without intending to be limited by any theory, the following examples are only to illustrate the products, preparation methods and uses of the present application, and are not intended to limit the scope of the invention of the present application.

Example

Example 1 Preparation of a nanometer-addressable fully functional DNA storage system

The design process of the nano-addressable DNA platform is as follows:

(1) The DNA origami skeleton chain uses M13mp18 single chain, and the staple chain combination is obtained according to the target template shape (two-dimensional rectangular structure). An exemplary manner may be to optionally mark the corresponding relationship between the staple chain and the address number according to the shape information of the seven-segment addressing structure. Randomly generate and screen out seven orthogonal address sequences with a length of 20 bases, and extend the staple chain sequences corresponding to addresses 1-7 respectively. The extended sequences are the designed address sequences, thereby obtaining the address sequence for assembly. A combination of staple strands that addresses the DNA platform. As shown in Figure 1, the information that can be stored in the seven data links corresponding to addresses 1-7 is the seven Chinese characters "Heaven and earth change and all things are connected".

(2) The switchable data link adopts a partially complementary paired double-stranded structure. A single Chinese character is converted into binary encoding and then encoded into a 10-base sequence using a cyclic encoding algorithm. The complete data chain is connected by the address complementary sequence, erasing functional region, T7 promoter sequence, and data sequence (including a pair of primers) (Figure 2). The closed strand contains 17 bases of the T7 promoter and the complementary sequence of 5 bases downstream and an 8-base dangling sequence corresponding to the address (called toehold). The data strand and closed strand form a double-stranded structure through partial complementary pairing of 22 bases.

Based on the above design, the structure of the nano-addressable platform is assembled. The assembly process is as follows:

(1) M13 single chain and addressable staple chain are mixed in 1×TAE-Mg ²⁺ solution, with a final concentration of 10 nM for the backbone chain and 50 nM for the staple chain.

(2) Perform annealing assembly in a PCR machine. The annealing procedure is: incubate at 95 degrees for 3 minutes, then cool to 25 degrees at a rate of 1 degree per minute, and finally maintain it at 4 degrees.

(3) The obtained assembly product is purified using PEG precipitation, and a pure assembly structure of approximately 20 nM is obtained as a substrate for information storage. For visualization purposes only, the obtained DNA structure can be characterized using atomic force microscopy (AFM).

(4) The data chain and the corresponding closed chain are mixed in 1×TAE-Mg ²⁺ solution, with a final concentration of 10 μM. Annealing and hybridization in a PCR machine. Hybridization products were characterized using 10% polyacrylamide gel electrophoresis (PAGE).

Embodiment 2 Data operation method of nanometer-addressable full-function DNA storage system

Data writing and reading:

(1) Add 100nM 1-7 data link to 10nM DNA origami, hybridize at room temperature for 1 hour, and write completely addressable information. At this time, the origami surface shows an "8" shape, consisting of 7*7 (49 in total) 1-7 data links. Use AFM to characterize the data writing success rate. Add some combinations of data chains 1-7 to form a "0-9" shape on the origami surface. For example, the number "1" is composed of two data chains, the number "2" is composed of 5 data chains, and the number "3" is composed of It consists of 5 data links, and so on. The robustness of data writing on the platform is visually verified through AFM morphology characterization. The results show (Figure 4) that the data chain of the target writing position can be visually verified in the AFM characterization results.

(2) Add the activation chains of data 1-7 to the solutions of data 1-7 respectively, react at room temperature for 1 hour, add the complete T7 promoter chain, and react at room temperature for 1 hour. Add T7 transcription mixture and react in a 42°C water bath for 1 hour. PAGE characterizes the viability of transcription upon activation and the orthogonality of addressable readouts. The results obtained by reading out the PAGE data statistics of the orthogonality of the activation (Figure 5) show that the data can only be exported if the sequence of the activation chain matches the data chain.

(3) The RNA molecules obtained by transcription are reverse transcribed using a reverse transcription kit to obtain the corresponding DNA strands. The obtained DNA strands are quantified using fluorescence quantitative PCR to confirm the export of the data.

(4) The exported data is sequenced using a second-generation sequencer and the sequencing results are decoded. The results of second-generation sequencing on the readout sequences of the seven address data showed that the sequence information obtained by sequencing at each address was completely consistent with the written information (Figure 6).

Addressable erasure and modification of data:

(1) After data links 1-7 are completely written, add erase chains 5 and 6 that are complementary to data links 5 and 6, incubate at room temperature for 1 hour, erase the corresponding data, and characterize the target data link through super-resolution fluorescence microscopy 5 and 6 have been erased, presenting the number "3". Further add more erasure chains 1, 4, and 7 that are complementary to data chains 1, 4, and 7 for reaction and characterization. Target data chains 1, 4, 7 is erased, resulting in the number "1". This process can be seen in Figure 7, where the number "8" is transformed into the number "3" and the number "1" in sequence.

(2) Add data link 1, incubate at room temperature for 1 hour, re-write data at address 1, and obtain the result of adding data link 1, showing the number "7". Through super-resolution fluorescence microscopy imaging results, the results show that addressable data can be erased and written. During each step of the process, the fluorescence imaging structural characterization results correspond to the target erase and write locations. This process can be seen in Figure 7, the transformation process from the number "1" to the number "7".

(3) Taking address 1 as an example to test the repeated modification ability, fix the DNA origami on the surface of the magnetic beads, and label the two data (two data chains) to be written to address 1 with Alexa488 and Cy5 fluorescent labels respectively. Add the Alexa488-labeled data link, use a fluorescence spectrophotometer to test the concentration of the data link remaining in the supernatant after writing, perform an erasure operation, and test the erasure efficiency. Write the Cy5-marked data link to test the writing efficiency, perform erasing, and test the erasing efficiency. The specific experiment is: first write 647 data, at this time the writing efficiency of the first 488 data is basically 0, then erase the 647 data, at this time the erasing efficiency of the 488 data is basically 0; in the next step, write the 488 data, At this time, the writing efficiency is about 54%, and then the 488 data is erased. At this time, the 488 data erasing efficiency is about 110%. And so on. Repeat more than 5 times to measure the addressable repeated erasing ability of the system. The results show that after repeated erasing and writing, the fluorescence test results obtained can correspond to the expected erasing and writing results, indicating that it is feasible to repeatedly modify the data (Figure 8). The upper part of the ordinate in Figure 8 represents the erasure of 488 data Efficiency, the bottom represents the writing efficiency of 488 data, and the abscissa represents the number of erases or writes.

Multiple erasing of data:

The 20 pM biotin-linked origami assembled in Example 1 was fixed on the glass slide through the PEG-biotin-Avidin method. After cleaning, 1 μM Cy5-labeled data link 1 was added, incubated for 30 minutes, washed, and fluorescence detection was performed under TIRF. Add 1 μM erase chain 1, incubate for 1 hour, wash with buffer, and take fluorescence photography again. The erasing and writing process was repeated 10 times, and TIRF (total internal reflection fluorescence microscope) recorded the fluorescence changes. Figure 9 is a single-point TIRF imaging diagram of seven address data being erased and written repeatedly for 10 times. Figure 9 shows that for a single fluorescent dot, it is very stable for 10 times of erasing. Figure 10 is a TIRF (total internal reflection fluorescence microscope) imaging diagram and co-localization statistical diagram of seven address data being repeatedly erased 10 times. The co-localization of red and green fluorescence represents the presence of data on the origami paper. Figure 10 shows that after 10 times of erasing and writing, the co-localization ratio after writing is stable at about 85%, and the co-localization ratio after erasing is stable at about 10%.

Multiple reads of data:

Assemble the data chain on the origami paper, add the activation chain for activation, adsorb 2nM origami paper on the mica sheet, clean it with 1×TAE Mg ²⁺ , add the transcription system on the mica, and incubate the mica in a 42°C oven for 2 hours Finally, the liquid on the mica is collected and stored. The above-mentioned transcription process was repeated 11 times, and the RNA transcribed each time was collected. Finally, the RNA transcribed 11 times was subjected to reverse transcription-PCR for quantification. Figure 11 is a quantitative PCR diagram of repeated readings of the data link. The results show that the storage medium of the present application can be read repeatedly for at least 10 times.

Example 3 Reading detection of origami platforms based on different matrix spacing

DNA origami is designed with different matrix spacings, including 6nm, 12nm, 18nm and 24nm, to store information chains Arrange them on the DNA origami paper at different spacings, give each information chain a physical address at different spacings, and detect the writing and reading of the data chains at different matrix spacings according to the method in Example 2. The results are shown in Figures 12 and 13. When the matrix spacing is divided into 6nm, 12nm, 18nm, and 24nm, there is no significant difference in the writing efficiency and readout efficiency of the data link.

The foregoing detailed description is provided by way of explanation and example, and is not intended to limit the scope of the appended claims. Various modifications to the embodiments described herein will be apparent to those of ordinary skill in the art and remain within the scope of the appended claims and their equivalents.

Claims (28)

1. A system comprising 1) a nucleic acid molecule containing data information, and 2) a carrier having addressable information, the nucleic acid molecule being able to bind to the carrier, and the nucleic acid molecule containing the data information being able to Reading is performed in situ on the vector.
2. The system of claim 1, wherein the carrier has different sequences of address sequences at different physical locations.
3. The system of claim 1, wherein the vector comprises a DNA origami substrate comprising a staple strand comprising an address sequence having addressable information.
4. The system of claim 3, wherein in the carrier, the address sequences of the staple chains are arranged in a matrix form.
5. The system of claim 4, wherein the distance between two adjacent staple chains is 6-24 nm.
6. The system of any one of claims 1-5, wherein the nucleic acid molecule comprises an address complementary sequence that is complementary to the address sequence on the vector.
7. The system of claim 6, wherein the address complementary sequence is about 15 or more nucleotides in length.
8. The system of any one of claims 1-7, wherein the nucleic acid molecule comprises a data sequence, and in a state where the nucleic acid molecule and the carrier are not substantially separated, the data information in the data sequence can is read.
9. The system of claim 8, wherein the data sequence is about 1 or more nucleotides in length.
10. The system of any one of claims 1-9, wherein the nucleic acid molecule includes a read priming sequence, the read priming sequence can trigger the synthesis of the data sequence into a strand to be sequenced, and the strand to be sequenced is identical to The data sequences are complementary.
11. The system of claim 10, wherein the read initiating sequence includes a promoter.
12. The system according to any one of claims 1-11, further comprising 3) a blocking chain, the blocking chain can prevent the data information from being read.
13. The system of claim 12, wherein the blocking strand includes a blocking sequence that is complementary to the read initiating sequence, thereby preventing the data information from being read.
14. The system of claim 13, wherein the blocking strand comprises a blocking elongation sequence, the blocking elongation sequence is located upstream and/or downstream of the blocking sequence, and the blocking elongation sequence is substantially non-complementary to the nucleic acid molecule. .
15. The system of any one of claims 12-14, further comprising 4) an activation chain, the activation chain can prevent the blocking chain from binding to the nucleic acid molecule, and the activation chain simultaneously binds to the nucleic acid molecule The blocking sequence and the blocking extension sequence are complementary.
16. The system of any one of claims 1-15, wherein the nucleic acid molecule comprises an erasure function sequence, the erasure function sequence is located upstream and/or downstream of the address complementary sequence, and the erasure function sequence The functional sequence is not substantially complementary to the address sequence on the carrier.
17. The system of claim 16, further comprising 5) an erasure chain, the erasure chain can prevent the nucleic acid molecule from binding to the carrier, and the erasure chain can simultaneously bind to the address complementary sequence and the erase and write function program Columns are complementary.
18. The system of claim 17, wherein the erasure strand is capable of displacing the carrier from binding to the nucleic acid molecule.
19. The system of claim 17, wherein the molar ratio of the nucleic acid molecule to the address sequence in the vector is 2:1 or higher.
20. The nucleic acid molecule in the system of any one of claims 1-19.
21. The vector in the system of any one of claims 1-19.
22. A method of data storage, the method comprising providing the system according to any one of claims 1-19.
23. A method of data editing, which method includes replacing data information in nucleic acid molecules in the system of any one of claims 1-19.
24. The method of claim 23, comprising providing the erasure strand, the nucleic acid molecule at a specific physical location binds to the erasure strand at room temperature, and the nucleic acid molecule is not substantially bound to the erasure strand. carrier combination.
25. A method of data reading, which method includes determining data information in nucleic acid molecules in the system of any one of claims 1-19.
26. The method of claim 25, further comprising providing the activation chain, at room temperature, the hindrance chain to which the nucleic acid molecule at a specific physical position is bound binds to the activation chain, and the hindrance chain is substantially Does not bind to the nucleic acid molecule.
27. A storage medium containing the method of any one of claims 22-26.
28. An apparatus comprising the storage medium of claim 27, and a processor coupled to the storage medium, the processor configured to execute based on a program stored in the storage medium to implement a right The method described in any one of claims 22-26.