CN107058300A - The tape label joint built for single-molecule sequencing library template and its application - Google Patents

The tape label joint built for single-molecule sequencing library template and its application Download PDF

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CN107058300A
CN107058300A CN201611195931.7A CN201611195931A CN107058300A CN 107058300 A CN107058300 A CN 107058300A CN 201611195931 A CN201611195931 A CN 201611195931A CN 107058300 A CN107058300 A CN 107058300A
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dna
sample
artificial sequence
tape label
sequence
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裴熙祥
孙孝政
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Harbin Botai Biotechnology Co Ltd
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    • C12Q1/6869—Methods for sequencing
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    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
    • C40B50/06—Biochemical methods, e.g. using enzymes or whole viable microorganisms

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Abstract

The invention belongs to Protocols in Molecular Biology and application field, and in particular to a kind of tape label joint built for single-molecule sequencing library template and its application, tape label joint sequence is as shown in numbering NO.1 ~ NO.96.The application process of label includes：The DNA that end-filling has been carried out after interrupting is used as original samples；To each sample, any one sequence is selected as the sample joint from the tape label joint sequence of 96 and not selected mistake；Reaction is attached to each sample and its corresponding joints；All sample mixed in equal amounts are set up into a sequencing library template.The sequencing library template of acquisition can be used for further operation using the sequencing of Pacbio microarray datasets.The present invention improves single-molecule sequencing platform sample flux, and effectively reduction single-molecule sequencing platform builds Kucheng's sheet, improves sequencing efficiency, is particularly suitable for use in several kb to the DNA sample to be measured of hundreds of Mb length, such as viral genome, BAC libraries, YAC libraries, plasmid.

Description

The tape label joint built for single-molecule sequencing library template and its application
Technical field
The invention belongs to biology field, and in particular to a kind of band mark built for single-molecule sequencing library template Sign joint and its application.
Background technology
By representative of Pacbio II microarray datasets the third generation single-molecule sequencing experiment in, before being sequenced, it is necessary to Sequencing library template is built, by the way that DNA sample is interrupted into certain length, then the DNA interrupted is repaired and end is mended The subsequent operations such as flat, adjunction head, purifying, complete library template and prepare.
Joint that usual adjunction head is used and without sequence label, each sequencing library template can only include a kind of sample Product, for DNA total lengths several kb to hundreds of Mb sample for example viral genome, BAC genomic libraries, YAC genomic libraries, Plasmid etc., the data volume that a sequencing library carries out a wheel sequencing generation is sequenced far beyond the target needed for obtaining desired result Amount, causes the sequencing wasting of resources.
By the way that target sequencing sequence is connected into DNA sequence tag, multiple samples can be mixed, be established as a survey Preface library template, by subsequent reactions, builds sequencing library, is sequenced.One sequencing reaction can be completed to multiple samples Sequencing, save sequencing resource, reduction sequencing cost.Tape label primer kit in the market is expensive, using by To serious limitation.
The content of the invention
The invention provides a kind of tape label joint built for single-molecule sequencing library template and its application, improve The sample flux of Pacbio microarray datasets once sequencing reaction, effectively reduction sequencing cost are DNA total lengths in several kb to hundreds of Mb sample such as viral genome, BAC genomic libraries, YAC genomic libraries, plasmid etc. provide tagging for high efficiency, low cost The method for building sequencing library template.
A kind of tape label joint for single-molecule sequencing library construction, for example following sequence number NO.1 of its base sequence ~ Shown in NO.96：
NO.1：5'pCATTCGCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGCGAATG3'；
NO.2：5'pCATCTCTGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCAGAGATG3'；
NO.3：5'pCATCCTAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTAGGATG3'；
NO.4：5'pCATGGATCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGATCCATG3'；
NO.5：5'pCATGCAAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTTGCATG3'；
NO.6：5'pCATATGGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCCATATG3'；
NO.7：5'pCATCTCGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCGAGATG3'；
NO.8：5'pCATGCTCGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGAGCATG3'；
NO.9：5'pCATACCAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTGGTATG3'；
NO.10：5'pCATCCGGTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATACCGGATG3'；
NO.11：5'pCATAACTCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGAGTTATG3'；
NO.12：5'pCATTTGAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTCAAATG3'；
NO.13：5'pCATACTATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATAGTATG3'；
NO.14：5'pCATTTGGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCCAAATG3'；
NO.15：5'pCATCGACCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGGTCGATG3'；
NO.16：5'pCATTAATGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCATTAATG3'；
NO.17：5'pCATAGGTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTACCTATG3'；
NO.18：5'pCATTGCGTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATACGCAATG3'；
NO.19：5'pCATGAATCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGATTCATG3'；
NO.20：5'pCATCATGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCATGATG3'；
NO.21：5'pCATACGCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGCGTATG3'；
NO.22：5'pCATGCATTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAATGCATG3'；
NO.23：5'pCATGATCTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGATCATG3'；
NO.24：5'pCATCAATAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTATTGATG3'；
NO.25：5'pCATTGACGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGTCAATG3'；
NO.26：5'pCATGATGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCATCATG3'；
NO.27：5'pCATCAATTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAATTGATG3'；
NO.28：5'pCATAGATAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTATCTATG3'；
NO.29：5'pCATCCGATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATCGGATG3'；
NO.30：5'pCATATGCCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGGCATATG3'；
NO.31：5'pCATCAGTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTACTGATG3'；
NO.32：5'pCATAATAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTATTATG3'；
NO.33：5'pCATCATCCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGGATGATG3'；
NO.34：5'pCATTCATGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCATGAATG3'；
NO.35：5'pCATAGAACATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGTTCTATG3'；
NO.36：5'pCATTGGAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTCCAATG3'；
NO.37：5'pCATCAGGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCCTGATG3'；
NO.38：5'pCATAATACATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGTATTATG3'；
NO.39：5'pCATCGAATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATTCGATG3'；
NO.40：5'pCATTTCGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCGAAATG3'；
NO.41：5'pCATAATTCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGAATTATG3'；
NO.42：5'pCATCGCGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCGCGATG3'；
NO.43：5'pCATAAGGTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATACCTTATG3'；
NO.44：5'pCATACTGGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCCAGTATG3'；
NO.45：5'pCATAGCAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTGCTATG3'；
NO.46：5'pCATGTACCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGGTACATG3'；
NO.47：5'pCATGGTCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGACCATG3'；
NO.48：5'pCATAATGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCATTATG3'；
NO.49：5'pCATAGTCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGACTATG3'；
NO.50：5'pCATAACTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTAGTTATG3'；
NO.51：5'pCATCTATGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCATAGATG3'；
NO.52：5'pCATCGACGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGTCGATG3'；
NO.53：5'pCATAACCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGGTTATG3'；
NO.54：5'pCATCGGCGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGCCGATG3'；
NO.55：5'pCATGCAGTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATACTGCATG3'；
NO.56：5'pCATCTCGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCGAGATG3'；
NO.57：5'pCATCTGCGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGCAGATG3'；
NO.58：5'pCATACGTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTACGTATG3'；
NO.59：5'pCATATACTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGTATATG3'；
NO.60：5'pCATTACTTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAAGTAATG3'；
NO.61：5'pCATAAGCTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGCTTATG3'；
NO.62：5'pCATGACGGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCCGTCATG3'；
NO.63：5'pCATAGAAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTTCTATG3'；
NO.64：5'pCATGTCCGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGGACATG3'；
NO.65：5'pCATTCAGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCTGAATG3'；
NO.66：5'pCATAGAGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCTCTATG3'；
NO.67：5'pCATGCCTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTAGGCATG3'；
NO.68：5'pCATTAATCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGATTAATG3'；
NO.69：5'pCATAACCTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGGTTATG3'；
NO.70：5'pCATGACGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCGTCATG3'；
NO.71：5'pCATTAGGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCCTAATG3'；
NO.72：5'pCATGGCATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATGCCATG3'；
NO.73：5'pCATTTCAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTGAAATG3'；
NO.74：5'pCATTTAACATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGTTAAATG3'；
NO.75：5'pCATTAGTCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGACTAATG3'；
NO.76：5'pCATTGCATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATGCAATG3'；
NO.77：5'pCATAATAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTATTATG3'；
NO.78：5'pCATAGCCTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGGCTATG3'；
NO.79：5'pCATCCAACATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGTTGGATG3'；
NO.80：5'pCATGCAGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCTGCATG3'；
NO.81：5'pCATAGAATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATTCTATG3'；
NO.82：5'pCATCAGCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGCTGATG3'；
NO.83：5'pCATTTCTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTAGAAATG3'；
NO.84：5'pCATCCTCTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGAGGATG3'
NO.85：5'pCATCCGGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCCGGATG3'；
NO.86：5'pCATGCCGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCGGCATG3'；
NO.87：5'pCATAACGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCGTTATG3'；
NO.88：5'pCATCCAGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCTGGATG3'；
NO.89：5'pCATTAGTTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAACTAATG3'；
NO.90：5'pCATTGGCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGCCAATG3'；
NO.91：5'pCATCGTATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATACGATG3'；
NO.92：5'pCATGCTAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTAGCATG3'；
NO.93：5'pCATGACTTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAAGTCATG3'；
NO.94：5'pCATGTACTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGTACATG3'；
NO.95：5'pCATCGAGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCTCGATG3'；
NO.96：5'pCATCGCAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTGCGATG3'.
Each sequence of tape label joint of the present invention is made up of with uniqueness label 5 ' ends and 3 ' the 8 pairs of complementary bases in end, Middle 45 bases are Pacbio microarray dataset universal linker sequences, and the sequence includes the sequence combined with sequencing primer.
Present invention also offers the template method that a kind of utilization tape label joint builds single-molecule sequencing library, including it is as follows Step：
（1）The DNA sample that end-filling has been carried out after interrupting is used as original samples；
（2）To each sample, any one sequence conduct is selected from the tape label joint sequence of 96 and not selected mistake The sample joint；
（3）Reaction is attached to each sample and its corresponding joints；
（4）All sample mixed in equal amounts are set up into a sequencing library template.
Step（1）The original samples are the DNA sample that end-filling has been carried out after interrupting, and the DNA sample can be But viral genome, BAC genomic libraries, YAC genomic libraries, plasmid etc. are not limited to for several kb treating to hundreds of Mb length Survey DNA sample.
Step（2）In to each sample, select any sequence from 96 tape label joint sequences shown in NO.1 ~ NO.96 Row are as jointing, and its selection principle is：Determine each original samples numbering, to each numbering sample, from NO.1 ~ A sequence is selected in NO.96, the sequence label of each sample is determined；In the optional NO.1 ~ NO.96 of sample each numbered It is any pair of, but the sample of different numberings can not select same tape label joint.
The tape label joint provided using the present invention has carried out the DNA sample difference of end-filling after can 96 be interrupted The different tape label joint of connection, then mixed in equal amounts is as a sequencing library template, for continuing storehouse and sequencing after progress, Greatly save and build storehouse and sequencing cost, improve and build storehouse and sequencing efficiency.
Step（3）The coupled reaction, reaction system is：
Overall reaction system is 20 μ L
Step（3）T4 DNA ligases and T4 DNA ligases buffer solution in its reaction system of the coupled reaction can be Takara Products, naturally it is also possible to be other company's like products.
Step（3）The coupled reaction, reaction condition is：22 DEG C of 30 min of connection, 65 DEG C, 5 min terminating reactions.
Step（4）It is described that all sample mixed in equal amounts are set up into a sequencing library template, it can be entered using Quibit 2.0 Row concentration mensuration, and all samples are subjected to mixed in equal amounts, naturally it is also possible to carry out concentration mensuration using other means such as Q-PCR Mixed in equal amounts afterwards.
The present invention builds single-molecule sequencing library template using tape label joint, is passed through after can disposably 96 be interrupted The different DNA samples of DNA end-fillings are mixed, and storehouse and sequencing are continued after being used for, and are greatlyd save reagent consumables cost, are carried The high sample flux of single sequencing reaction.Compared with the tape label joint that Pacbio companies provide, same company can be reached Efficiency is connect, but reagent cost is substantially reduced.Tape label joint of the present invention can be used for all based on Pacbio single-molecule sequencing platforms DNA library is built.
Brief description of the drawings
Fig. 1 is the mixing sequencing result overview of 7 samples.
Embodiment
For the object of the invention, technical scheme and advantage is expanded on further, with reference to the preferred embodiments of the present invention to this Explanation is explained in detail, but the present invention is not limited in the range of the present embodiment.
Unreceipted particular technique condition in embodiment, according to the technology bar described by document or product description in the art Part is carried out.Agents useful for same or the unreceipted production firm of instrument, be this area conventional products.
Embodiment 1
The DNA sample that end-filling has been carried out after interrupting is used as original samples：
Herpesviral is double-stranded DNA virus, and size is about 140 kb, will interrupt 7 plants of blebs for 3-10 kb by end-filling Viral DNA is used as the original samples for building sequencing library.
Embodiment 2
Implement by taking above-mentioned 7 samples as an example
It is 1 to 7 by 7 sample number consecutivelies, a sequence is each chosen from NO.1 ~ NO.96 bar tape label joint sequences As the sample joint sequence, sample and joint corresponding table are made
Embodiment 3
Reaction is attached to above-mentioned 7 samples and its corresponding joints
Coupled reaction system is：
Overall reaction system is 20 μ L
Coupled reaction condition is：22 DEG C of 30 min of connection, 65 DEG C, 5 min terminating reactions.
Embodiment 4
All sample mixed in equal amounts are set up into a sequencing library template
7 samples after connection are respectively taken into 10 μ L, a new 0.2 μ L centrifugation is transferred to and is in control for subsequently continuing to build storehouse The mixing sequencing library template containing 7 samples of sequencing, is directly used in next step and builds storehouse sequencing or be placed in -20 DEG C of refrigerators It is standby.
SEQUENCE LISTING
<110>Harbin Bo Tai bio tech ltd
<120>The tape label joint built for single-molecule sequencing library template and its application
<130>
<160> 96
<170> PatentIn version 3.3
<210> 1
<211> 61
<212> DNA
<213>Artificial sequence
<400> 1
cattcgcaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattgcgaat 60
g 61
<210> 2
<211> 61
<212> DNA
<213>Artificial sequence
<400> 2
catctctgat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcagagat 60
g 61
<210> 3
<211> 61
<212> DNA
<213>Artificial sequence
<400> 3
catcctagat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatctaggat 60
g 61
<210> 4
<211> 61
<212> DNA
<213>Artificial sequence
<400> 4
catggatcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgatccat 60
g 61
<210> 5
<211> 61
<212> DNA
<213>Artificial sequence
<400> 5
catgcaagat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcttgcat 60
g 61
<210> 6
<211> 61
<212> DNA
<213>Artificial sequence
<400> 6
catatggaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattccatat 60
g 61
<210> 7
<211> 61
<212> DNA
<213>Artificial sequence
<400> 7
catctcgaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattcgagat 60
g 61
<210> 8
<211> 61
<212> DNA
<213>Artificial sequence
<400> 8
catgctcgat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcgagcat 60
g 61
<210> 9
<211> 61
<212> DNA
<213>Artificial sequence
<400> 9
cataccaaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatttggtat 60
g 61
<210> 10
<211> 61
<212> DNA
<213>Artificial sequence
<400> 10
catccggtat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gataccggat 60
g 61
<210> 11
<211> 61
<212> DNA
<213>Artificial sequence
<400> 11
cataactcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgagttat 60
g 61
<210> 12
<211> 61
<212> DNA
<213>Artificial sequence
<400> 12
catttgaaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatttcaaat 60
g 61
<210> 13
<211> 61
<212> DNA
<213>Artificial sequence
<400> 13
catactatat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatatagtat 60
g 61
<210> 14
<211> 61
<212> DNA
<213>Artificial sequence
<400> 14
catttggaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattccaaat 60
g 61
<210> 15
<211> 61
<212> DNA
<213>Artificial sequence
<400> 15
catcgaccat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatggtcgat 60
g 61
<210> 16
<211> 61
<212> DNA
<213>Artificial sequence
<400> 16
cattaatgat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcattaat 60
g 61
<210> 17
<211> 61
<212> DNA
<213>Artificial sequence
<400> 17
cataggtaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattacctat 60
g 61
<210> 18
<211> 61
<212> DNA
<213>Artificial sequence
<400> 18
cattgcgtat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatacgcaat 60
g 61
<210> 19
<211> 61
<212> DNA
<213>Artificial sequence
<400> 19
catgaatcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgattcat 60
g 61
<210> 20
<211> 61
<212> DNA
<213>Artificial sequence
<400> 20
catcatgcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgcatgat 60
g 61
<210> 21
<211> 61
<212> DNA
<213>Artificial sequence
<400> 21
catacgcaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattgcgtat 60
g 61
<210> 22
<211> 61
<212> DNA
<213>Artificial sequence
<400> 22
catgcattat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gataatgcat 60
g 61
<210> 23
<211> 61
<212> DNA
<213>Artificial sequence
<400> 23
catgatctat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatagatcat 60
g 61
<210> 24
<211> 61
<212> DNA
<213>Artificial sequence
<400> 24
catcaataat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattattgat 60
g 61
<210> 25
<211> 61
<212> DNA
<213>Artificial sequence
<400> 25
cattgacgat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcgtcaat 60
g 61
<210> 26
<211> 61
<212> DNA
<213>Artificial sequence
<400> 26
catgatgcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgcatcat 60
g 61
<210> 27
<211> 61
<212> DNA
<213>Artificial sequence
<400> 27
catcaattat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gataattgat 60
g 61
<210> 28
<211> 61
<212> DNA
<213>Artificial sequence
<400> 28
catagataat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattatctat 60
g 61
<210> 29
<211> 61
<212> DNA
<213>Artificial sequence
<400> 29
catccgatat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatatcggat 60
g 61
<210> 30
<211> 61
<212> DNA
<213>Artificial sequence
<400> 30
catatgccat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatggcatat 60
g 61
<210> 31
<211> 61
<212> DNA
<213>Artificial sequence
<400> 31
catcagtaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattactgat 60
g 61
<210> 32
<211> 61
<212> DNA
<213>Artificial sequence
<400> 32
cataatagat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatctattat 60
g 61
<210> 33
<211> 61
<212> DNA
<213>Artificial sequence
<400> 33
catcatccat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatggatgat 60
g 61
<210> 34
<211> 61
<212> DNA
<213>Artificial sequence
<400> 34
cattcatgat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcatgaat 60
g 61
<210> 35
<211> 61
<212> DNA
<213>Artificial sequence
<400> 35
catagaacat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgttctat 60
g 61
<210> 36
<211> 61
<212> DNA
<213>Artificial sequence
<400> 36
cattggaaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatttccaat 60
g 61
<210> 37
<211> 61
<212> DNA
<213>Artificial sequence
<400> 37
catcaggaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattcctgat 60
g 61
<210> 38
<211> 61
<212> DNA
<213>Artificial sequence
<400> 38
cataatacat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgtattat 60
g 61
<210> 39
<211> 61
<212> DNA
<213>Artificial sequence
<400> 39
catcgaatat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatattcgat 60
g 61
<210> 40
<211> 61
<212> DNA
<213>Artificial sequence
<400> 40
catttcgcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgcgaaat 60
g 61
<210> 41
<211> 61
<212> DNA
<213>Artificial sequence
<400> 41
cataattcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgaattat 60
g 61
<210> 42
<211> 61
<212> DNA
<213>Artificial sequence
<400> 42
catcgcgcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgcgcgat 60
g 61
<210> 43
<211> 61
<212> DNA
<213>Artificial sequence
<400> 43
cataaggtat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gataccttat 60
g 61
<210> 44
<211> 61
<212> DNA
<213>Artificial sequence
<400> 44
catactggat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatccagtat 60
g 61
<210> 45
<211> 61
<212> DNA
<213>Artificial sequence
<400> 45
catagcagat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatctgctat 60
g 61
<210> 46
<211> 61
<212> DNA
<213>Artificial sequence
<400> 46
catgtaccat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatggtacat 60
g 61
<210> 47
<211> 61
<212> DNA
<213>Artificial sequence
<400> 47
catggtcaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattgaccat 60
g 61
<210> 48
<211> 61
<212> DNA
<213>Artificial sequence
<400> 48
cataatgaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattcattat 60
g 61
<210> 49
<211> 61
<212> DNA
<213>Artificial sequence
<400> 49
catagtcaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattgactat 60
g 61
<210> 50
<211> 61
<212> DNA
<213>Artificial sequence
<400> 50
cataactaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattagttat 60
g 61
<210> 51
<211> 61
<212> DNA
<213>Artificial sequence
<400> 51
catctatgat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcatagat 60
g 61
<210> 52
<211> 61
<212> DNA
<213>Artificial sequence
<400> 52
catcgacgat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcgtcgat 60
g 61
<210> 53
<211> 61
<212> DNA
<213>Artificial sequence
<400> 53
cataaccaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattggttat 60
g 61
<210> 54
<211> 61
<212> DNA
<213>Artificial sequence
<400> 54
catcggcgat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcgccgat 60
g 61
<210> 55
<211> 61
<212> DNA
<213>Artificial sequence
<400> 55
catgcagtat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatactgcat 60
g 61
<210> 56
<211> 61
<212> DNA
<213>Artificial sequence
<400> 56
catctcgcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgcgagat 60
g 61
<210> 57
<211> 61
<212> DNA
<213>Artificial sequence
<400> 57
catctgcgat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcgcagat 60
g 61
<210> 58
<211> 61
<212> DNA
<213>Artificial sequence
<400> 58
catacgtaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattacgtat 60
g 61
<210> 59
<211> 61
<212> DNA
<213>Artificial sequence
<400> 59
catatactat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatagtatat 60
g 61
<210> 60
<211> 61
<212> DNA
<213>Artificial sequence
<400> 60
cattacttat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gataagtaat 60
g 61
<210> 61
<211> 61
<212> DNA
<213>Artificial sequence
<400> 61
cataagctat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatagcttat 60
g 61
<210> 62
<211> 61
<212> DNA
<213>Artificial sequence
<400> 62
catgacggat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatccgtcat 60
g 61
<210> 63
<211> 61
<212> DNA
<213>Artificial sequence
<400> 63
catagaagat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcttctat 60
g 61
<210> 64
<211> 61
<212> DNA
<213>Artificial sequence
<400> 64
catgtccgat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatcggacat 60
g 61
<210> 65
<211> 61
<212> DNA
<213>Artificial sequence
<400> 65
cattcagcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgctgaat 60
g 61
<210> 66
<211> 61
<212> DNA
<213>Artificial sequence
<400> 66
catagagcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgctctat 60
g 61
<210> 67
<211> 61
<212> DNA
<213>Artificial sequence
<400> 67
catgcctaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattaggcat 60
g 61
<210> 68
<211> 61
<212> DNA
<213>Artificial sequence
<400> 68
cattaatcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgattaat 60
g 61
<210> 69
<211> 61
<212> DNA
<213>Artificial sequence
<400> 69
cataacctat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gataggttat 60
g 61
<210> 70
<211> 61
<212> DNA
<213>Artificial sequence
<400> 70
catgacgaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattcgtcat 60
g 61
<210> 71
<211> 61
<212> DNA
<213>Artificial sequence
<400> 71
cattaggcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgcctaat 60
g 61
<210> 72
<211> 61
<212> DNA
<213>Artificial sequence
<400> 72
catggcatat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatatgccat 60
g 61
<210> 73
<211> 61
<212> DNA
<213>Artificial sequence
<400> 73
catttcaaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatttgaaat 60
g 61
<210> 74
<211> 61
<212> DNA
<213>Artificial sequence
<400> 74
catttaacat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgttaaat 60
g 61
<210> 75
<211> 61
<212> DNA
<213>Artificial sequence
<400> 75
cattagtcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgactaat 60
g 61
<210> 76
<211> 61
<212> DNA
<213>Artificial sequence
<400> 76
cattgcatat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatatgcaat 60
g 61
<210> 77
<211> 61
<212> DNA
<213>Artificial sequence
<400> 77
cataataaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatttattat 60
g 61
<210> 78
<211> 61
<212> DNA
<213>Artificial sequence
<400> 78
catagcctat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gataggctat 60
g 61
<210> 79
<211> 61
<212> DNA
<213>Artificial sequence
<400> 79
catccaacat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgttggat 60
g 61
<210> 80
<211> 61
<212> DNA
<213>Artificial sequence
<400> 80
catgcagaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattctgcat 60
g 61
<210> 81
<211> 61
<212> DNA
<213>Artificial sequence
<400> 81
catagaatat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatattctat 60
g 61
<210> 82
<211> 61
<212> DNA
<213>Artificial sequence
<400> 82
catcagcaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattgctgat 60
g 61
<210> 83
<211> 61
<212> DNA
<213>Artificial sequence
<400> 83
catttctaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattagaaat 60
g 61
<210> 84
<211> 61
<212> DNA
<213>Artificial sequence
<400> 84
catcctctat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatagaggat 60
g 61
<210> 85
<211> 61
<212> DNA
<213>Artificial sequence
<400> 85
catccggaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattccggat 60
g 61
<210> 86
<211> 61
<212> DNA
<213>Artificial sequence
<400> 86
catgccgcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgcggcat 60
g 61
<210> 87
<211> 61
<212> DNA
<213>Artificial sequence
<400> 87
cataacgaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattcgttat 60
g 61
<210> 88
<211> 61
<212> DNA
<213>Artificial sequence
<400> 88
catccagcat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatgctggat 60
g 61
<210> 89
<211> 61
<212> DNA
<213>Artificial sequence
<400> 89
cattagttat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gataactaat 60
g 61
<210> 90
<211> 61
<212> DNA
<213>Artificial sequence
<400> 90
cattggcaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattgccaat 60
g 61
<210> 91
<211> 61
<212> DNA
<213>Artificial sequence
<400> 91
catcgtatat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatatacgat 60
g 61
<210> 92
<211> 61
<212> DNA
<213>Artificial sequence
<400> 92
catgctaaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatttagcat 60
g 61
<210> 93
<211> 61
<212> DNA
<213>Artificial sequence
<400> 93
catgacttat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gataagtcat 60
g 61
<210> 94
<211> 61
<212> DNA
<213>Artificial sequence
<400> 94
catgtactat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatagtacat 60
g 61
<210> 95
<211> 61
<212> DNA
<213>Artificial sequence
<400> 95
catcgagaat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gattctcgat 60
g 61
<210> 96
<211> 61
<212> DNA
<213>Artificial sequence
<400> 96
catcgcagat ctctctcttt tcctcctcct ccgttgttgt tgttgagaga gatctgcgat 60
g 61

Claims (5)

1. a kind of tape label joint for single-molecule sequencing library construction, it is characterised in that its base sequence such as one sequence Shown in numbering NO.1 ~ NO.96：
NO.1：5'pCATTCGCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGCGAATG3'；
NO.2：5'pCATCTCTGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCAGAGATG3'；
NO.3：5'pCATCCTAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTAGGATG3'；
NO.4：5'pCATGGATCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGATCCATG3'；
NO.5：5'pCATGCAAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTTGCATG3'；
NO.6：5'pCATATGGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCCATATG3'；
NO.7：5'pCATCTCGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCGAGATG3'；
NO.8：5'pCATGCTCGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGAGCATG3'；
NO.9：5'pCATACCAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTGGTATG3'；
NO.10：5'pCATCCGGTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATACCGGATG3'；
NO.11：5'pCATAACTCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGAGTTATG3'；
NO.12：5'pCATTTGAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTCAAATG3'；
NO.13：5'pCATACTATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATAGTATG3'；
NO.14：5'pCATTTGGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCCAAATG3'；
NO.15：5'pCATCGACCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGGTCGATG3'；
NO.16：5'pCATTAATGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCATTAATG3'；
NO.17：5'pCATAGGTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTACCTATG3'；
NO.18：5'pCATTGCGTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATACGCAATG3'；
NO.19：5'pCATGAATCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGATTCATG3'；
NO.20：5'pCATCATGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCATGATG3'；
NO.21：5'pCATACGCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGCGTATG3'；
NO.22：5'pCATGCATTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAATGCATG3'；
NO.23：5'pCATGATCTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGATCATG3'；
NO.24：5'pCATCAATAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTATTGATG3'；
NO.25：5'pCATTGACGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGTCAATG3'；
NO.26：5'pCATGATGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCATCATG3'；
NO.27：5'pCATCAATTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAATTGATG3'；
NO.28：5'pCATAGATAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTATCTATG3'；
NO.29：5'pCATCCGATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATCGGATG3'；
NO.30：5'pCATATGCCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGGCATATG3'；
NO.31：5'pCATCAGTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTACTGATG3'；
NO.32：5'pCATAATAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTATTATG3'；
NO.33：5'pCATCATCCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGGATGATG3'；
NO.34：5'pCATTCATGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCATGAATG3'；
NO.35：5'pCATAGAACATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGTTCTATG3'；
NO.36：5'pCATTGGAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTCCAATG3'；
NO.37：5'pCATCAGGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCCTGATG3'；
NO.38：5'pCATAATACATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGTATTATG3'；
NO.39：5'pCATCGAATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATTCGATG3'；
NO.40：5'pCATTTCGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCGAAATG3'；
NO.41：5'pCATAATTCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGAATTATG3'；
NO.42：5'pCATCGCGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCGCGATG3'；
NO.43：5'pCATAAGGTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATACCTTATG3'；
NO.44：5'pCATACTGGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCCAGTATG3'；
NO.45：5'pCATAGCAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTGCTATG3'；
NO.46：5'pCATGTACCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGGTACATG3'；
NO.47：5'pCATGGTCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGACCATG3'；
NO.48：5'pCATAATGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCATTATG3'；
NO.49：5'pCATAGTCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGACTATG3'
NO.50：5'pCATAACTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTAGTTATG3'；
NO.51：5'pCATCTATGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCATAGATG3'；
NO.52：5'pCATCGACGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGTCGATG3'；
NO.53：5'pCATAACCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGGTTATG3'；
NO.54：5'pCATCGGCGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGCCGATG3'；
NO.55：5'pCATGCAGTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATACTGCATG3'；
NO.56：5'pCATCTCGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCGAGATG3'；
NO.57：5'pCATCTGCGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGCAGATG3'；
NO.58：5'pCATACGTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTACGTATG3'；
NO.59：5'pCATATACTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGTATATG3'；
NO.60：5'pCATTACTTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAAGTAATG3'；
NO.61：5'pCATAAGCTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGCTTATG3'；
NO.62：5'pCATGACGGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCCGTCATG3'；
NO.63：5'pCATAGAAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTTCTATG3'；
NO.64：5'pCATGTCCGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCGGACATG3'；
NO.65：5'pCATTCAGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCTGAATG3'；
NO.66：5'pCATAGAGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCTCTATG3'；
NO.67：5'pCATGCCTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTAGGCATG3'；
NO.68：5'pCATTAATCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGATTAATG3'；
NO.69：5'pCATAACCTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGGTTATG3'；
NO.70：5'pCATGACGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCGTCATG3'；
NO.71：5'pCATTAGGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCCTAATG3'；
NO.72：5'pCATGGCATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATGCCATG3'；
NO.73：5'pCATTTCAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTGAAATG3'；
NO.74：5'pCATTTAACATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGTTAAATG3'；
NO.75：5'pCATTAGTCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGACTAATG3'；
NO.76：5'pCATTGCATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATGCAATG3'；
NO.77：5'pCATAATAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTATTATG3'；
NO.78：5'pCATAGCCTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGGCTATG3'；
NO.79：5'pCATCCAACATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGTTGGATG3'；
NO.80：5'pCATGCAGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCTGCATG3'；
NO.81：5'pCATAGAATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATTCTATG3'；
NO.82：5'pCATCAGCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGCTGATG3'；
NO.83：5'pCATTTCTAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTAGAAATG3'；
NO.84：5'pCATCCTCTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGAGGATG3'；
NO.85：5'pCATCCGGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCCGGATG3'；
NO.86：5'pCATGCCGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCGGCATG3'；
NO.87：5'pCATAACGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCGTTATG3'；
NO.88：5'pCATCCAGCATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATGCTGGATG3'；
NO.89：5'pCATTAGTTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAACTAATG3'
NO.90：5'pCATTGGCAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTGCCAATG3'；
NO.91：5'pCATCGTATATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATATACGATG3'；
NO.92：5'pCATGCTAAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTTAGCATG3'；
NO.93：5'pCATGACTTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAAGTCATG3'；
NO.94：5'pCATGTACTATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATAGTACATG3'；
NO.95：5'pCATCGAGAATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATTCTCGATG3'；
NO.96：5'pCATCGCAGATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGATCTGCGATG3'.
2. tape label joint according to claim 1 builds the template method in single-molecule sequencing library, it is characterised in that bag Include following steps：
（1）The DNA sample that end-filling has been carried out after interrupting is used as original samples；
（2）To each sample, any one sequence conduct is selected from the tape label joint sequence of 96 and not selected mistake The sample joint；
（3）Reaction is attached to each sample and its corresponding joints；
（4）All sample mixed in equal amounts are set up into a sequencing library template.
3. method according to claim 2, it is characterised in that the sample can be, but not limited to, viral genome, BAC Genomic library, YAC genomic libraries, plasmid etc. are to be measured DNA samples of several kb to hundreds of Mb length.
4. coupled reaction according to claim 2, reaction system is：
Herpesviral end-filling reaction product 8 μL(84 ng) T4 DNA ligases 1 μL(350 U) T4 DNA ligase buffer solutions 2 μL 20 μM of joints 1 5 μL Deionized water 4 μL
Reaction system is 20 μ L.
5. coupled reaction according to claim 2, reaction condition is：22 DEG C of 30 min of connection, 65 DEG C, 5 min terminate anti- Should.
CN201611195931.7A 2016-12-22 2016-12-22 The tape label joint built for single-molecule sequencing library template and its application Pending CN107058300A (en)

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CN109136222A (en) * 2018-09-13 2019-01-04 武汉菲沙基因信息有限公司 The tape label connector of PacBio microarray dataset Multi-example mixing sequencing library building and application
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