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

Establishment method of gene library of fecal flora based on high-throughput gene sequencing

A fecal flora and gene sequencing technology, applied in the biological field, can solve the problems of low throughput, low efficiency, and cannot well meet the needs of scientific research and clinical practice, and achieve the effect of fast library construction and large throughput.

Inactive Publication Date: 2016-09-14
GUANGZHOU SAGENE BIOTECH
View PDF4 Cites 26 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In traditional molecular biology methods, the Sanger method is sometimes used to sequence and identify 16SrDNA of a single strain, but limited by low throughput and low efficiency, it cannot well meet the needs of scientific research and clinical practice

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
  • Establishment method of gene library of fecal flora based on high-throughput gene sequencing
  • Establishment method of gene library of fecal flora based on high-throughput gene sequencing
  • Establishment method of gene library of fecal flora based on high-throughput gene sequencing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0113] The technical solutions of the present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments, but the present invention is not limited to the following embodiments. The materials used in the examples are the feces of a group of patients confirmed to be positive for Clostridium difficile AB toxin provided by the General Hospital of Guangzhou Military Region.

[0114] The specific steps of the experimental process for establishing the library of the present invention are as follows.

[0115] 1. Extract the total genomic DNA of fecal flora:

[0116]The total genomic DNA of the fecal flora was extracted strictly according to the operation manual of the TIANamp Stool DNA Kit (TIANGEN DP328), and the conditions were optimized for intestinal bacteria: the lysis temperature of the sample pretreatment was adjusted to 95°C and incubated for 10 minutes, so that the comparative Difficult-to-break Gram-positive ba...

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

An establishment method of a gene library of fecal flora based on high-throughput gene sequencing employs a ''nested PCR'' method for enrichment amplification on 16S rDNA, so as to reduce the host and food residue genome pollution by the maximum; and V3 and V6 are combined for specific amplification and massive parallel sequencing, so as to obtain the a target gene sequence database of the flora. The library can increase the identification of the bacterial flora from Genus level to Species level. The method can maximally avoid the interference of host cell nucleic acid, and completely and efficiently detect 16SrDNA tag sequence of all bacteria varieties, so as to accurately determine the ecological structure of the intestinal flora.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection methods for bacteria in the field of biotechnology, and in particular relates to a method for establishing a fecal flora gene library based on high-throughput gene sequencing. Background technique [0002] In recent years, the relationship between gut microbes, diet and health has been increasingly explained, largely due to the rapid development of gut microbiome research technology. The research on gut microbes has generally gone through three stages: culture-dependent methods, culture-independent traditional molecular biology methods, and sequencing-based high-throughput omics methods. [0003] By comparing the characteristics and applications of the research methods in the three stages, people can find that the culture-dependent method can obtain the corresponding strains while identifying the strains, which is convenient for subsequent research, but the culture method itsel...

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): C40B50/06C12N15/10C12Q1/68C12Q1/04
Inventor 伍泳彰
Owner GUANGZHOU SAGENE BIOTECH
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