Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

BRCA (breast cancer susceptibility gene) high-throughput sequencing library and construction method and application thereof

A construction method and sequencing library technology, which are applied in the field of BRCA high-throughput sequencing library and its construction, can solve the problems of loss of target fragments, cross-contamination, and excessive steps, and achieve automatic construction, improved utilization, and simple operation. Effect

Pending Publication Date: 2019-08-16
安徽鼎晶生物科技有限公司 +1
View PDF10 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The classic high-throughput library construction method is currently the most widely accepted library construction method with the most stable data performance and the least data bias, but the library construction process takes a long time, there are too many steps and the loss of target fragments during the purification process , so that the template utilization rate of the library is not high, and may cause cross-contamination
The classic high-throughput library construction method is increasingly unable to meet the needs of the fast-growing market for low-input micro-library construction and fast reporting cycles

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
  • BRCA (breast cancer susceptibility gene) high-throughput sequencing library and construction method and application thereof
  • BRCA (breast cancer susceptibility gene) high-throughput sequencing library and construction method and application thereof
  • BRCA (breast cancer susceptibility gene) high-throughput sequencing library and construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] A method for constructing a BRCA high-throughput sequencing library, comprising generating an initial library, purifying the initial library, generating a sequencing library, and purifying the sequencing library.

[0076] Step 1, generation of the initial library: Multiplex PCR is used to capture and amplify the target fragments. For the specific process, add 0.025ml of reaction system into a 0.2ml PCR tube, and perform amplification according to specific reaction procedures. The specific reaction system and procedures are shown in Table 1 and Table 2.

[0077] Table 1. Reaction system of initial library

[0078] Element volume Polymerase Mix 12.5μl Primer library 7μl template 0.5μl (total 10ng) water 5μl

[0079] Table 2. Initial library amplification program

[0080]

[0081] Step 2. Purification of the initial library: use magnetic beads to purify to remove primers and replace the reaction solution, so as to perform A ...

Embodiment 2

[0101] Automated build solution

[0102] In the first step, inject 1 μl template into the bottom of the target well of the 96-well plate, then place the 96-well plate on the 96-well plate rack, extend the probe into the top of the 96-well plate, switch the channel to pipeline A so that the pipeline is filled with the first round of PCR For the reaction solution, open the channel, and inject 29 μl of the first-round PCR reaction solution into the target well. Remove the 96-well plate, paste the sealing film, shake and mix well, centrifuge and put it on the PCR instrument for reaction. The components of the reaction solution are shown in Table 1, and the reaction conditions are shown in Table 2.

[0103] In the second step, switch the pipeline to pipeline B so that the pipeline is filled with the magnetic bead mixture, take out the 96-well plate after the first round of PCR from the PCR instrument and place it on the plate rack, and insert the probe into the top of the PCR tube...

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 invention provides a construction method of a BRCA (breast cancer susceptibility gene) high-throughput sequencing library and belongs to the field of high-throughput sequencing. The construction method provided herein comprises generating an initial library, purifying the initial library, generating a sequencing library and purifying the sequencing library. The whole construction method provided herein is performed in a vessel, so that cross infection probabilities are avoided. The construction method provided herein allows purification to be efficient and convenient since magnetic bead reaction is combined with washing operation. The construction method provided herein allows utilization rate of a template to be increased greatly.

Description

technical field [0001] The invention belongs to the technical field of high-throughput sequencing, and in particular relates to a BRCA high-throughput sequencing library and its construction method and application. Background technique [0002] Breast cancer susceptibility genes (BRCA1 / 2) were first discovered in breast cancer families, and are susceptibility genes for breast and ovarian cancers with genetic predisposition. BRCA1 is located on human chromosome 17 q21 and is inherited in an autosomal dominant manner with high penetrance. BRCA1 is about 100kb long and contains 24 exons, and its gene product is a phosphorylated protein composed of 1863 amino acids. BRCA2 is located on human chromosome 13 q12, the whole genome DNA is about 70kb in length, and the coding region is about 10kp in length. The gene sequence of BRCA2 has no obvious relationship with BRCA1. It consists of 27 exons, of which the 11th exon is about 4932bp in length, the mRNA is about 10.2kb in length, ...

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): C12Q1/6806C12N15/10C40B50/06
CPCC12Q1/6806C12N15/1093C40B50/06C12Q2531/113C12Q2537/143C12Q2523/308C12Q2535/122
Inventor 沈伟强胡文玮焦海涛何志辉熊江红谢秒秒张瑞华叶嘉惠李飞燕朱建华廖敔方姝葛海鹏
Owner 安徽鼎晶生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com