Target region enrichment sequencing method for 165 genes

A target region and gene technology, applied in the field of genetic engineering and biotechnology detection, can solve problems such as high cost

Inactive Publication Date: 2017-02-22
刘鹏飞
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In addition to whole genome sequencing and whole exome sequencing, the

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  • Target region enrichment sequencing method for 165 genes
  • Target region enrichment sequencing method for 165 genes
  • Target region enrichment sequencing method for 165 genes

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Embodiment Construction

[0102] The present invention and its beneficial technical effects are described in detail below in conjunction with various specific embodiments, wherein: this embodiment is based on the technical solution, and provides detailed implementation and specific operation process, but the protection scope of the present invention does not limited to this example.

[0103] 1. Use the Qiagen ctDNA extraction kit to extract the free DNA in the plasma of 5 different samples, quantify the DNA and dissolve it in 50ul ddH 2 In O, library construction was performed. The required reagent preparations are listed in Table 1.

[0104] Reagent name storage method preparation method Buffer ACL* room temperature room temperature Buffer ACW1 room temperature room temperature Buffer ACW2 room temperature room temperature Buffer ACB room temperature room temperature Proteinase K -20℃ on ice Buffer AVE room temperature room temperature...

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Abstract

The invention provides a target region enrichment sequencing method for 165 genes. According to the method, 165 group-entering genes are screened according to the cancer genome and medicine data of TCGA, NCCN, FDA and the like. The 165 genes comprise detection genes recommended by NCCN, target genes corresponding to target medicines and important clinical medicines approved by FDA, genes influencing the medication effect of the target medicines, and genes influencing the chemical drug sensibility and toxic and side effects. According to the method, the exon regions of the 165 genes are enriched at once through the probe capturing technology, the multi-gene and multi-target parallel depth high-throughput sequencing is realized, the various variation types (such as point mutation, deletion, insertion, fusion and copy number amplification) of the genes can be simultaneously detected, and the detection depth and accuracy are superior to those of the prior art.

Description

technical field [0001] The invention relates to the field of genetic engineering and biotechnology detection, in particular to a method for enriching target regions of 165 genes, which is used for multi-target parallel high-throughput sequencing of circulating tumor DNA (ctDNA), and simultaneously checks multiple genes Variation types, such as mutations, deletions, insertions, fusions, and copy number amplifications, outperform existing technologies in depth and breadth of detection. Background technique [0002] Circulating DNA is a kind of extracellular DNA in a cell-free state, which exists in body fluids such as blood, synovial fluid, and cerebrospinal fluid. It is mainly composed of single-stranded or double-stranded DNA and a mixture of single-stranded and double-stranded DNA. Complex or free DNA exists in two forms. As a new tumor marker, circulating DNA will play an important role in the diagnosis, treatment and prognosis of tumors, especially for some tumors that d...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/10
CPCC12N15/1013C12Q1/6869C12Q1/6886C12Q2535/122
Inventor 刘鹏飞张文会朱东兴韩雪
Owner 刘鹏飞
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