Construction method and applications of tumor BRCA1/2 gene variation library for high-throughput sequencing detection

A technology of gene mutation and construction method, which is applied to the construction of tumor BRCA1/2 gene mutation library for high-throughput sequencing detection and its application field, which can solve the problems of high clinical charges, low sensitivity and low throughput, etc.

Pending Publication Date: 2017-11-03
XIAMEN SPACEGEN BIOTECH CO LTD
View PDF4 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Existing tumor gene detection mainly uses fluorescent quantitative PCR method, which has a low detection throughput. For the whole exome detection of BRCA1 and BRCA2 genes, the cost is high and the sample size is large, resulting in high clinical fees and insufficient sample size.
At present, the

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Construction method and applications of tumor BRCA1/2 gene variation library for high-throughput sequencing detection
  • Construction method and applications of tumor BRCA1/2 gene variation library for high-throughput sequencing detection
  • Construction method and applications of tumor BRCA1/2 gene variation library for high-throughput sequencing detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] A method for constructing a tumor BRCA1 / 2 gene mutation library for high-throughput sequencing detection, covering all exon region mutations of the human BRCA1 / 2 gene. The specific exon mutation information is shown in Table 1:

[0039] Table 1

[0040]

[0041]

[0042]

[0043]

[0044]

[0045] Specifically include the following steps:

[0046] A method for constructing a tumor BRCA1 / 2 gene variation library for high-throughput sequencing detection, covering all exon region mutations of the human BRCA1 / 2 gene, specifically comprising the following steps:

[0047](1) Design the first basic amplification primer set and the second basic amplification primer set for the target gene BRCA1 / 2, all forward primers and reverse primers of the first basic amplification primer set and the second basic amplification primer set The 5' ends of the primers are equipped with additional 2-5 Ts, and the first T of the 2-5 Ts near the 3' end has PNA modification, and the...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The present invention discloses a construction method of a tumor BRCA1/2 gene variation library for high-throughput sequencing detection, wherein the tumor BRCA1/2 gene variation library covers the mutations at all the exon regions of the human BRCA1/2 gene. According to the present invention, the complete exon sequences of the BRCA1 gene and the BRCA2 gene are subjected to single-tube amplification to rapidly complete the construction of the library, wherein the construction time of the whole library is only 2-3 h, and the manual time is only 15-30 min; and by combining the obtained library and the high-throughput sequencing platform, the difficult problem that the detection of the complete exon sequence is required on the basis of the small amount of the clinical samples in tumor diseases in the clinic can be effectively solved, and the cost is low.

Description

technical field [0001] The invention specifically relates to a method for constructing a tumor BRCA1 / 2 gene variation library for high-throughput sequencing detection and its application. Background technique [0002] Breast cancer and ovarian cancer are the most common malignant tumors that threaten women's health. The incidence of breast cancer in women varies significantly in different countries, with the highest incidence in the United States and Northern Europe. The occurrence and development of breast cancer are related to many factors, which are the result of the joint action of environmental factors and genetic factors. BRCA1 and BRCA2 are the abbreviations of Breast Cancer Susceptibility Gene1 and 2, that is, breast cancer susceptibility gene 1 / 2. Among them, the BRCA1 gene is located on human chromosome 17q21 and consists of 23 exons; the BRCA2 gene is located on human chromosome 13q12.3 and consists of 27 exons. Both BRCA1 and BRCA2 genes are tumor suppressor g...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/10C12Q1/68C40B50/06
CPCC12N15/1093C12Q1/6806C40B50/06C12Q2525/107C12Q2525/173C12Q2531/113
Inventor 陈琰郭飞飞
Owner XIAMEN SPACEGEN BIOTECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap