Unlock instant, AI-driven research and patent intelligence for your innovation.
Pretreatment method for extracting metagenome DNA of intestinal contents
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A technology of metagenomic and content, which is applied in the field of pretreatment for the extraction of metagenomic DNA from intestinal content, and achieves the effect of strong practicability
Active Publication Date: 2017-12-15
广东美格基因科技有限公司
View PDF2 Cites 5 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
[0004] The purpose of the present invention is to: overcome the problems that the existing method is difficult to extract metagenomic DNA in the intestinal substance of corrosive fish, and provide a pretreatment method for extracting the metagenomic DNA of intestinal contents. After conventional DNA extraction steps, the complete metagenomic DNA of intestinal contents can be extracted efficiently
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
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0030] The method of the present invention extracts the metagenomic DNA of the intestinal contents of bighead carp:
[0031] (1) Take 0.25g of fresh bighead carp intestinal content sample into a 15ml centrifuge tube, add a decongestant, fully vortex, and centrifuge at 11,000g for 10min (can be washed twice).
[0032] (2) Discard the supernatant, add a mixture of detergent + 0.5mM EDTA, and vortex well.
[0033] (3) Freeze at -80°C for 15 minutes, and heat at 65°C for 5 minutes (repeated 3 times).
[0043] The method of the present invention extracts the metagenomic DNA of the intestinal contents of grass carp:
[0044] (1) Take 0.25g of fresh grass carp intestinal content sample into a 15ml centrifuge tube, add a decontamination agent, fully vortex, and centrifuge at 11,000g for 10min (can be washed twice).
[0045] (2) Discard the supernatant, add a mixture of detergent + 1 mM EDTA, and vortex well.
[0046] (3) Freeze at -80°C for 15 minutes, and heat at 65°C for 5 minutes (repeated 3 times).
[0048](5) Add proteinase K, 10 molarmass % SDS at 65° C., 1 h (upside down every 15 min).
[0049] (6) RNase was added at 37° C. for 15 min (to a final concentration of 20 mg / ml).
[0050] (7) Centrifuge at 11000rpm for 10min at 4°C.
[0051] (8) Pipette the supernatant into a new 50ml centrifuge tube, add equal volume of phenol:chloroform:isoamyl alcohol (25:24:1) and extract twice (the protein layer can be seen)...
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
Login to View More
Abstract
The invention discloses a pretreatment method for extracting metagenome DNA of intestinal contents. The pretreatment method comprises the following steps of: adding a necrotic-tissue removing agent into the intestinal contents to wash for twice, then adding nuclease eliminating solution for washing, cracking cells by adopting a freezing-thawing method and a cracking method, fully releasing DNA in the cells, finally adding protease K, SDS and RNAenzyme to remove protein, fiber and RNA in the cells, and treating products by adopting a conventional method to obtain the metagenome DNA of the intestinal contents. The pretreatment method disclosed by the invention has the advantages that the effect for extracting the DNA of the metagenome DNA of the intestinal contents of omnivorous animals is obvious, and both the extracting purity and the extracting amount can meet downstream experiments such as PCR augmentation and high-throughput sequencing and the like, so that the practicability is very strong.
Description
technical field [0001] The invention belongs to the technical field of gene extraction, and more particularly relates to a pretreatment method for the extraction of metagenomic DNA from intestinal contents. Background technique [0002] Fish gut microbes play a very important role in fish development and health. In 1998, the American Academy of Sciences first proposed the concept of metagenomics. By analyzing the metagenome of fish gut contents, the usual dietary habits of fish can be known. The composition of probiotics / harmful bacteria and other foreign invading bacteria in the gut, so as to better understand the balance of the intestinal environment of fish and the composition of bacteria. However, due to the extremely complex environment of fish in water and many food sources for fish, serious degradation or failure to extract occurs during the extraction process. Extraction is very difficult. [0003] At present, the methods of extracting fish gut metagenomes include ...
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
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to View More 
Login to View More