Capture probe for DNA sample containing INDEL region, kit and library construction method

A capture probe and library construction technology, applied in the field of high-throughput sequencing, can solve problems such as low capture efficiency of target fragments, and achieve the effect of solving low capture efficiency and improving capture specificity and sensitivity

Pending Publication Date: 2019-12-13
BEIJING USCI MEDICAL LAB CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The main purpose of the present invention is to provide a capture probe, kit and library construction method for DNA samples containi

Method used

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  • Capture probe for DNA sample containing INDEL region, kit and library construction method
  • Capture probe for DNA sample containing INDEL region, kit and library construction method
  • Capture probe for DNA sample containing INDEL region, kit and library construction method

Examples

Experimental program
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Effect test

Embodiment 2

[0078] Through the IDT online probe design software (Target Capture Probe Design&Ordering Tool), using the human reference genome Human (Feb.2009GRCh37 / hg19) as the reference genome, a probe covering the entire exon region of the complete EGFR gene was designed, excluding the UTR interval. The length of each probe is 120nt, and the multiplier is 2×. The EGFR gene has a total of 28 exons of different lengths, and the designed shingled single-stranded DNA probe sequence ensures complete coverage of each region. For chromosome 20 insertion of EGFR gene, the insertion sequence is underlined. Two types of panels (panels) are formed by designing probes with traditional IDT software and the probe design scheme of the present invention respectively, and the specific design schemes are as follows.

[0079] Conventional hybridization capture single-stranded DNA probe design. Use the IDT online probe design software to design imbricate single-stranded DNA probe sequences for the mutati...

Embodiment 3

[0089] Through the IDT online probe design software (Target Capture Probe Design&Ordering Tool), using the human reference genome Human (Feb.2009GRCh37 / hg19) as the reference genome, a probe covering the entire exon region of the complete EGFR gene was designed, excluding the UTR interval. For the BRCA2 gene short fragment deletion A1689 deletion, the base sequence of the deletion is GCA. Two types of panels (panels) are formed by designing probes with traditional IDT software and the probe design scheme of the present invention respectively, and the specific design schemes are as follows.

[0090] Conventional hybridization capture single-stranded DNA probe design. Use IDT online probe design software to design shingled single-stranded DNA probe sequences for BRCA2 gene mutation sites, as shown in Table 13 below, where SEQ ID NO13~SEQ ID NO15 shows that each probe is 120bp in length, and the overlapping interval The 5' end of the probe is labeled with biotin. Each probe is m...

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Abstract

The invention provides a capture probe for a DNA sample containing an INDEL region, a kit and a library construction method. The capture probe comprises a first section of probe sequence which is complementarily paired with the left side of the INDEL region, a second section of probe sequence which is complementarily paired with the right side of the INDEL region, and a third section of probe sequence which is completely complementarily paired with the INDEL region. According to Indel (insertion-deletion fragment) of a target region, a mutated gene sequence is used as a reference sequence, andthree probes are designed for the Indel region and are respectively an Indel left side probe sequence, an Indel right side probe sequence and a probe sequence covering an Indel mutation site. The capture specificity and sensitivity of the three probes to the target Indel sequence are greatly improved, and the problems of low capture efficiency and low enrichment level of the Indel region of an important gene at present are effectively solved.

Description

technical field [0001] The invention relates to the technical field of high-throughput sequencing, in particular to a capture probe, a kit and a library construction method for a DNA sample containing an INDEL region. Background technique [0002] With the completion of the Human Genome Project, human beings have made unprecedented progress in understanding and mastering their own genetic information. At the same time, the continuous development and improvement of the genetic testing technology platform at the molecular level has led to the rapid development of genetic testing technology and the continuous improvement of genetic testing efficiency. Next-generation sequencing (NGS) technology has the advantages of large throughput, short time consumption, high accuracy and rich information, and can accurately locate genes in a short time, mainly including whole genome sequencing (WGS), whole Exome sequencing (whole exome sequencing, WES) and targeted sequencing (targeted reg...

Claims

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Application Information

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IPC IPC(8): C12Q1/6827C12N15/11C40B40/06C40B50/06
CPCC12Q1/6827C40B40/06C40B50/06
Inventor 方楠王燕刘运超伍启熹王建伟刘倩唐宇
Owner BEIJING USCI MEDICAL LAB CO LTD
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