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Multiple PCR primer group and kit for detecting gene related to colorectal cancer administration

A colorectal cancer, gene detection technology, applied in recombinant DNA technology, biochemical equipment and methods, microbial determination/inspection, etc. The effect of starting volume reduction

Active Publication Date: 2017-11-28
GUANGZHOU FOREVERGEN BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, hybridization capture is characterized by the ability to capture the exome or even a larger target region, but the operation process is more complicated and requires special equipment; the multiplex PCR operation is simple and flexible, and only needs a PCR machine. Complete target sequence enrichment and library construction within hours, suitable for relatively small target sequence enrichment

Method used

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  • Multiple PCR primer group and kit for detecting gene related to colorectal cancer administration
  • Multiple PCR primer group and kit for detecting gene related to colorectal cancer administration
  • Multiple PCR primer group and kit for detecting gene related to colorectal cancer administration

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Design and synthesis of multiplex PCR primers for colorectal cancer individualized drug gene mutation detection

[0039] Use the primer design software whose patent application title is a multiplex PCR primer and its design method for amplifying BRCA1 / 2 gene (application number is 201610737156.7) to design primers, and input the genomic region (bed file format, per The line content is "chromosome-tab-start coordinate-tab-end coordinate"), and the output is multiple pairs of primers designed to be amplified in one reaction. The specific implementation ideas are as follows: 1) Use Primer3 software to design as many available primers as possible for each target region as alternatives; 2) According to the specificity of primer amplification, whether the primer contains simple repeat sequences, and whether the primer contains High-frequency SNP sites (defined as sites with a minor allele frequency of more than five thousandths in dbSNP) and other conditions filter ...

Embodiment 2

[0043] Example 2 Different types of sample library construction and mutation detection

[0044] 1. Experimental materials

[0045] The samples involved in this test are: genomic DNA of normal human blood, genomic DNA of MCF10A cell line, and FFPE genomic DNA of colorectal cancer. Genomic DNA of various types of samples was extracted using DNA extraction kits.

[0046] Genomic DNA extraction: use Qiagen DNeasy Blood&Tissue Kit, and follow the kit instructions to extract genomic DNA. The obtained genomic DNA is subjected to gel electrophoresis and quality detection using NanoDrop100. It is required that the genomic DNA has no obvious degradation, the concentration is greater than 20ng / μL, A260 / A280>1.8, A260 / A230>2.0, and Qubit 3.0 is used for accurate quantification.

[0047] A detection kit for colorectal cancer drug-related genes, including a primer set in Example 1, a universal downstream primer (URS), and a linker primer for Illuminate sequencer sequencing library construc...

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Abstract

The invention discloses a multiple PCR primer group for detecting a gene related to colorectal cancer administration. The primer group comprises an upstream primer group and a downstream primer group, wherein the 5' end sequence of the upstream is complementary to partial sequence of a to-be-detected primer, and the 3' end of the upstream primer is a specific sequence combined to a target zone; the 5' end sequence of the downstream primer is complementary with partial sequence of the to-be-detected primer, and the 3' end of the downstream primer is a specific sequence combined to the target zone, and the sequence of the middle position of the downstream primer is a molecule tag sequence. The invention further discloses a kit containing the multiple PCR primer group and a method for detecting the gene related to colorectal cancer administration, which can be used for simultaneously detecting exon regions of 8 genes related to target medicines for treating colorectal cancer and mutation of 23 polymorphic sites related to chemotherapeutic drugs, can be directly used for assisting clinical colorectal cancer administration, and can be used in cancer early diagnosis, auxiliary diagnosis and screening or cancer prognosis monitoring.

Description

technical field [0001] The invention relates to the technical field of dihydrate gypsum preparation, in particular to a multiplex PCR primer set, kit and detection method for colorectal cancer individualized drug-related gene detection. Background technique [0002] Colorectal cancer (CRC) is a general term for colon cancer and rectal cancer. higher. In addition to surgical resection and radiation therapy, current cancer treatment includes chemotherapy and targeted drug therapy targeting cancer cells. Targeted drug therapy uses unique structures or molecules in cancer cells to attack these special structures with specific drugs to kill cancer cells. Most importantly, targeted drugs are less likely to attack normal cells and cause less damage to normal cells, which can reduce many side effects (such as white blood cells, thrombocytopenia, anemia, vomiting, hair loss, etc.), and can also inhibit the growth of cancer cells, Malignant transformation capacity for metastasis an...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6886C12Q2600/106C12Q2600/118C12Q2600/156C12Q2537/143
Inventor 赖炳权何广良唐毅张纪斌李伟琴林钊许少飞
Owner GUANGZHOU FOREVERGEN BIOTECH CO LTD
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