Adapter for next-generation sequencing

A second-generation sequencing and sequencing technology, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as unrecognized and hindered research fields

Active Publication Date: 2017-07-11
北京迈基诺基因科技股份有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0002] The error rate of next-generation sequencing technology itself is about 1%, which is acceptable for some applications (such as genetic disease-causing genes, SNP site detection, etc.), but for metagenomics, paleontological genomics, cancer, etc. The field is a very big obstacle. These samples invol...

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  • Adapter for next-generation sequencing
  • Adapter for next-generation sequencing
  • Adapter for next-generation sequencing

Examples

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Embodiment 1

[0083] Example 1. A complete set of reagents used as adapters can improve the accuracy of high-throughput sequencing

[0084] This embodiment provides a kit of reagents for next-generation sequencing as an adapter, which consists of two single-stranded DNAs named A strand and B strand;

[0085] The A chain is shown in formula (I) from the 5' end to the 3' end:

[0086] A1-A3-A2(I);

[0087] Each nucleotide of A3 is A, T, C or G, that is, A3 is a random sequence;

[0088] The B chain is shown in formula (II) from the 3' end to the 5' end:

[0089] B1-B2(II);

[0090] A2 and B2 are complementary; A1 and B1 are not complementary;

[0091] A3 is not complementary to both B1 and A1;

[0092] The sequences of A1 and A2 are different; the sequences of B1 and B2 are different;

[0093] The 3' terminal nucleotide in A2 is a thionucleotide, that is, the oxygen atom with a double bond on the phosphoric acid in the 3' terminal nucleotide is replaced by a sulfur atom; the 5' terminal...

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Abstract

The invention discloses an adapter for next-generation sequencing. The adapter disclosed in the invention is composed of a single-stranded DNA A and a single-stranded DNA B or composed of a single-stranded DNA C and a single-stranded DNA D. The single-stranded DNA A, from terminal 5' to terminal 3', is as shown in a formula (I), i.e., A1-A3-A2, wherein each nucleotide of A3 is A, T, C or G; the single-stranded DNA B, from terminal 3' to terminal 5', is as shown in a formula (II), i.e., B1-B2, wherein A2 and B2 are complementary, A1 and B1 are not complementary, A3, B1 and A1 are not complementary, the sequences of A1 and A2 are different, and the sequences of B1 and B2 are different; the single-stranded DNA C is composed of A1 and A2; and the single-stranded DNA D is composed of B1, B2 and A3. Experimental results show that the adapter provided by the invention can easily and efficiently reduce false positive mutation in next-generation sequencing so as to realize more sensitive detection of low-frequency mutation in heterogeneous mixed samples like heterogeneous samples and chimera samples of tumors.

Description

technical field [0001] The invention relates to an adapter used for next-generation sequencing in the field of biotechnology. Background technique [0002] The error rate of next-generation sequencing technology itself is about 1%, which is acceptable for some applications (such as genetic disease-causing genes, SNP site detection, etc.), but for metagenomics, paleontological genomics, cancer, etc. The field is a very big obstacle. These samples involving deep sequencing and complex heterogeneity need to detect less than 1% of rare mutations, and 1% of background mutations in next-generation sequencing make less than 1% of rare mutations unrecognizable, so there is an urgent need for a More accurate sequencing methods meet the needs of current sequencing. Contents of the invention [0003] The technical problem to be solved by the present invention is how to improve the accuracy of sequencing. [0004] In order to solve the above technical problems, the present invention...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6806C12Q1/6869C12Q2525/191C12Q2531/113C12Q2535/122
Inventor 伍建姬晓雯
Owner 北京迈基诺基因科技股份有限公司
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