Linker sequence and method for detecting ultra-low frequency mutation of target sequence

A linker sequence and target sequence technology, which is applied in the field of linker sequences with ultra-low frequency mutations, to achieve the effect of reducing the dosage and solving the heterogeneity of tumor cells

Inactive Publication Date: 2017-12-08
IGENETECH BIOTECH (BEIJING) CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to solve the problem of ultra-low frequency mutation detection, the inventors ...

Method used

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  • Linker sequence and method for detecting ultra-low frequency mutation of target sequence
  • Linker sequence and method for detecting ultra-low frequency mutation of target sequence
  • Linker sequence and method for detecting ultra-low frequency mutation of target sequence

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Embodiment

[0053] In the embodiment of the present invention, a certain amount of tagged molecules are obtained by performing end-repair, A addition, adapter ligation, fragment enrichment and capture on the cfDNA standard HD780, which is then sequenced on the computer, combined with The later bioanalytical method corrects the positive and negative chains and analyzes the real mutation information.

[0054] Design and synthesis of linker sequences with molecular tags:

[0055] MID1:

[0056] AATGATACGGCGACCACCGAGATCTACACNNNNNNNNACACTCTTTCCCTACACGACGCTCTTCCGATCTAGATTGACT (SEQ ID NO. 1)

[0057] MID-1:

[0058] GTCAATCTAGATCGGAAGCAGCACACGTCTGAACTCCAGTCACAATGATGAATCTCGTATGCCGTCTTCTGCTTG (SEQ ID NO. 2)

[0059] MID2:

[0060] AATGATACGGCGACCACCGAGATCTACACNNNNNNNNACACTCTTTCCCTACACGACGCTCTTCCGATCTTACGTGACT (SEQ ID NO. 3)

[0061] MID2-2:

[0062] GTCAGCTAAGATCGGAAGACACACGTCTGAACTCCAGTCACAATGATGAATCTCGTATGCCGTCTTCTGCTTG (SEQ ID NO. 4)

[0063] MID3:

[0064] AATGATACGGCGACCACCGAGATCTACAC...

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Abstract

The invention discloses a linker sequence and method for detecting the ultra-low frequency mutation of a target sequence. According to the method, a certain number of labeled molecules can be obtained by conducting terminal repair, A adding, linker connection, fragment enrichment, capture and the like on an interrupted genome or cfDNA, then on-machine sequencing is conducted, positive and negative chains are corrected by means of a later-period biological analysis method, and real mutation information is obtained through analysis.

Description

technical field [0001] The invention belongs to the field of high-throughput sequencing, and in particular relates to a linker sequence and a method for detecting ultra-low frequency mutation of a target sequence. Background technique [0002] Compared with the first-generation sequencing technology, the second-generation sequencing technology can detect tens of billions of sequence information at a time with less time and expense. It is possible to enter the era of precision medicine. At present, due to the advantages of high throughput, high efficiency, and moderate price of next-generation sequencing technology, it has been widely used in tumor detection, highlighting its advantages. However, during library construction, the high-fidelity enzyme has a replication error rate of 10 -6 , the error rate will also increase with the increase of the number of cycles. At the same time, during the sequencing process, the reading rate of the bases by the instrument is between 0.0...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q1/6869C12Q2600/156C12Q2535/122C12Q2525/191
Inventor 余越美王瑞超邵谦之屈武斌蔡万世
Owner IGENETECH BIOTECH (BEIJING) CO LTD
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