Extraction method for metagenome

A metagenome and extraction method technology, applied in the field of metagenome extraction, can solve problems such as multi-time, and achieve the effects of increasing output, reducing waste of sequencing data, and reducing high background interference

Inactive Publication Date: 2014-12-17
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION +1
View PDF1 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For infectious disease samples, the method of dealing with DNA contamination takes more time, or requires special handling methods when collecting samples

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
  • Extraction method for metagenome
  • Extraction method for metagenome
  • Extraction method for metagenome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1 Probe hybridization subtraction for simulated mixed samples (hybridization first and then coupling)

[0071] 1. Take 20 μl of simulated mixed DNA (according to human whole blood DNA and 2815bp plasmid mixed according to the nanogram amount of 9:1, the total amount is about 100ng / μl) in a 200 μl tube, and place it in a PCR instrument to preheat at 95°C for 5 minutes. 65°C for 5 minutes; take 27 μl of 2×hybridization solution HYB and place it in a PCR instrument to preheat at 65°C for 5 minutes;

[0072] 2. Take 0.1 μl each of the probes whose serial numbers are SEQ ID NO: 1~70, about 70 pmol, and preheat at 65°C for 2 minutes;

[0073] 3. Mix 54 μl of the above system, hybridize at 65°C for 1 hour;

[0074] 4. Take 30 μl of magnetic beads M-280 (invitrogen), wash twice with 2×B&W buffer, discard the supernatant, and resuspend with 54 μl 2×B&W;

[0075] 5. Add the hybridization system (54 μl) to the magnetic beads, mix well, and incubate at room temperature ...

Embodiment 2

[0079] Example 2 Probe hybridization subtraction for total DNA in respiratory lavage fluid samples (hybridization first and then coupling)

[0080] Except in Step 1 of Example 1, select human respiratory lavage fluid sample DNA, take 32 μl of 2×hybridization solution HYB, and take 3 μl (about 120 pmol) of each probe with sequence number SEQ ID NO: 1~4 in Step 2, Except for resuspending with 64 μl 2×B&W in step 4, the same method and conditions as in Example 1 were used to carry out first hybridization followed by coupled probe hybridization subtraction.

Embodiment 3

[0081] Example 3 Probe hybridization subtraction for simulated mixed samples (coupling first and then hybridization)

[0082] 1. Take 30μl magnetic beads M-280 (invitrogen), wash twice with 2×B&W buffer, discard the supernatant, add 15μl 2×B&W buffer to resuspend the magnetic beads;

[0083] 2. Add 0.1 μl (about 70 pmol) and 8 μl of ddH to the resuspended magnetic beads 2 O, incubate at room temperature for 15 minutes, and mix gently during the period; place on a magnetic stand at room temperature for 1 to 2 minutes, and remove the supernatant;

[0084] 3. Take 20 μl of simulated mixed DNA (the human whole blood DNA and about 2.6kbp plasmid are mixed according to the nanogram amount of 9:1, the total amount is about 100ng / μl) in a 200μl PCR tube, and put in the PCR machine to preheat at 95°C for 5 minutes , 65°C for 5 minutes; take 26 μl of 2×hybridization solution HYB and place it in a PCR instrument to preheat at 65°C for 5 minutes;

[0085] 4. Add the mixed DNA and hybr...

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 an extraction method for metagenome, and a method of probe hybridization-bead capture is used to reduce the host DNA content in a host metagenomic DNA sample. According to the method, the Alu repeat sequence of the host DNA sequences and conserved sequence at both ends thereof are used as templates to design one-way or two-way probes, wherein the 5 'end of each probe is modified with biotin, streptavidin coated beads are used to capture host DNA hybridized with probes, in order to achieve aims of weakening host DNA background in a metagenome, and enhancing sequencing data efficiency.

Description

technical field [0001] The invention relates to a method for extracting a metagenomic group, in particular to a method for reducing host DNA in a host metagenomic DNA sample. Background technique [0002] A metagenome is all genetic material present in an environmental sample, including the genomes of many individual species. Metagenomics refers to the science of obtaining genetic material directly from environmental samples and conducting research on metagenomics. With the development of high-throughput DNA sequencing and bioinformatics, the application value of metagenomics continues to increase. By analyzing the metagenomic library, the population characteristics and interaction rules of microorganisms in an environment can be found, and the interaction between microorganisms and the environment can be explored. By sequencing the DNA of microbial populations, genes with specific functions can be screened to obtain proteins with special functions. [0003] The human res...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 任鲁风王绪敏楚亚男高静谷岚隋硕康禹徐力袁丽娜张奕杰高占成于军
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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