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

Construction method of RNA library for enriching original transcript information and application thereof

A construction method and transcript technology, applied in the field of RNA library construction, can solve the problems of lack of original transcript information, sequencing information results, etc.

Active Publication Date: 2016-04-20
BGI SHENZHEN CO LTD
View PDF1 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But using this method, the obtained mRNA does not contain many other poly(A)-transcripts that can be used for transcriptional analysis
Moreover, the loss of original transcript information often requires additional cycles of amplification reactions for small amounts of RNA input, and even this results in loss of some original transcript information and suboptimal sequencing information

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 of RNA library for enriching original transcript information and application thereof
  • Construction method of RNA library for enriching original transcript information and application thereof
  • Construction method of RNA library for enriching original transcript information and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] The construction of embodiment 1 RNA library

[0087] Specific experimental steps (see figure 1 Process steps shown in ):

[0088] 1. RNaseH purification of total RNA

[0089] 1.1 Take 1ug or 3ug of total RNA and anneal to oligoDNA

[0090]

[0091] 1.2 React at 95°C for 2 minutes on a PCR machine; cool down gradually, from 0.1°C to 22°C every 1 second; react at 22°C for 5 minutes, and place on ice quickly.

[0092] 1.3 RNaseH enzyme digestion

[0093]

[0094] React at 37°C for 30min

[0095] 1.4 DNaseI enzyme digestion

[0096]

[0097] 1.5 After the reaction, purify with RNAcleanXP magnetic beads and dissolve in 10ul nuclease-free water.

[0098] 2. RNA Fragmentation

[0099] Add 3 μL of 5×first-strand buffer to the eluate in the previous step, at 94°C for 10 min, and place on ice immediately.

[0100] 3. Reverse Transcribed First-Strand Synthesis

[0101] 3.1 Add 0.5 μL N6primer (0.1 μg / μL) to the RNA obtained in the previous step.

[0102] 3.2 Re...

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 high-quality RNA library for enriching original transcript information by using RNase H and selectively removing lots of rRNA and blood globulin and an application thereof. The RNA library constructed by the method has very low rRNA residual volume (which is only 1 / 500 of residual volume in a conventional oligo(dT) method), and has very high gene coverage.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to a method for constructing an RNA library enriched with original transcript information and its application. Background technique [0002] With the application and development of high-throughput sequencing technology, RNA sequencing technology is becoming more and more mature. Usually, when we construct RNA libraries, especially RNA-seq libraries, transcriptome libraries, and strand-specific transcription profile libraries, we need to perform mRNA purification on TotalRNA. Since the most abundant RNA in human cells is rRNA, accounting for about 82% of the total RNA, and the proportion of mRNA is only 4%. At present, the oligod(T) method is used to capture mRNA, relying on oligo(dT) to separate poly(A )+RNA to achieve the purpose of screening out a large amount of rRNA to obtain purified mRNA. But using this method, the obtained mRNA does not contain many other...

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): C40B50/06C40B40/08
Inventor 纪晓钧郭晶祝珍珍耿春雨李计广章文蔚蒋慧
Owner BGI SHENZHEN CO LTD
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