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A kind of multiplex PCR primer for amplifying brca1/2 gene and a kind of design method of multiplex PCR primer

A design method and primer design technology, applied in biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc., can solve problems such as complex operation procedures, achieve high quality, simple operation, and uniform amplification sexual effect

Active Publication Date: 2019-11-08
GUANGZHOU FOREVERGEN HEALTH TECH CO LTD +1
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  • 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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  • A kind of multiplex PCR primer for amplifying brca1/2 gene and a kind of design method of multiplex PCR primer
  • A kind of multiplex PCR primer for amplifying brca1/2 gene and a kind of design method of multiplex PCR primer
  • A kind of multiplex PCR primer for amplifying brca1/2 gene and a kind of design method of multiplex PCR primer

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Embodiment 1

[0034] Concrete implementation process of the present invention is as follows:

[0035] 1. Multiplex PCR primer design software development and hereditary breast cancer BRCA1 / 2 primer design.

[0036] The input of the multiplex PCR primer design software is the genomic region that needs to be designed for primers (bed file format, the content of each line is "chromosome-tab-start coordinates-tab-end coordinates"), and the output is designed and available in Multiple pairs of primers for amplification 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, Filter the primers designed in step 1 based on whether the primers contain simple repeat sequences, whether there are high-frequency SNP sites on the primers (defined as sites with a minor allele frequency of more than 5 / 1000 in dbSNP) and other con...

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Abstract

The invention provides a multiplex PCR primer for amplifying a BRCA1 / 2 gene and a design method of the multiplex PCR primer. The multiplex PCR primer comprises nucleotide sequences shown by SEQ ID NO:1 to SEQ ID NO:234. The invention further provides the design method of the multiplex PCR primer. With the multiplex PCR primer provided by the invention, the enrichment of the BRCA1 / 2 target sequence can be rapidly completed, further, the NGS library construction and sequencing are carried out, and then the highly-automatic and high-throughput detection for the BRCA1 / 2 gene of the hereditary breast cancer is realized.

Description

technical field [0001] The invention relates to a sequence of multiple PCR primers, in particular to the application of multiple PCR primers for amplifying BRCA1 / 2 genes and a design method thereof in genetic detection of hereditary breast cancer. Background technique [0002] Breast cancer is the cancer with the highest incidence of malignant tumors among women in my country. According to the data of the World Health Organization (WHO), in 2008, the incidence of breast cancer in Chinese women was 21.6 / 100,000, with 169,000 new cases and 45,000 deaths. According to existing research reports, about 5% to 10% of breast cancers are caused by BRCA1 / 2 gene mutations. BRCA1 / 2 mutations account for a large proportion (40% to 45%) of hereditary breast cancer patients, and 80% of patients in breast cancer high-risk families have mutations in the BRCA1 / 2 gene. BRCA1 mutation carriers have a lifetime risk of breast cancer of more than 80%, and BRCA2 mutation carriers have a lifetime ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11C40B40/08
CPCC12Q1/6886C12Q2600/156C12Q2600/16C40B40/08
Inventor 张纪斌王辉云林钊李伟琴罗景燕赖炳权
Owner GUANGZHOU FOREVERGEN HEALTH TECH CO LTD
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