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

A method for breaking the wall of filamentous fungal genome dna extraction

A technology for filamentous fungi and genomes, which is applied in the field of wall breaking necessary for the extraction of filamentous fungal genomes, can solve problems such as high cost, environmental protection hazards of chemical reagents, and long time consumption, so as to reduce dependence, reduce bacterial pollution, and facilitate operation Effect

Inactive Publication Date: 2016-01-06
CHINA THREE GORGES UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chemical reagents have hidden dangers in environmental protection
[0003] The methods developed to obtain high-quality DNA are generally time-consuming, expensive, and require some special equipment, which is difficult for general biological laboratories to meet.

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] A simple wall-breaking method for extracting filamentous fungal genome DNA, the method comprising the following steps:

[0016] 1) Filamentous fungus culture: culture filamentous fungus A in NBRIP [National Botanical Research Institute’sphosphatemedium (pH7.0), containing: 10gglucose, 5gCa per liter 3 (PO 4 ) 2 ,5gMgCl 2 ·6H 2 O,0.25gMgSO 4 ·7H 2 O, 0.2gKCl, 0.1g (NH 4 ) 2 SO 4 ] medium, a 250ml Erlenmeyer flask was loaded with 50ml liquid medium, cultured with shaking at 28°C for 2 days, and then cultured statically for 4 days;

[0017] 2) Collect fungal mycelium: Use a sterilized toothpick to pick the fungus A cultured in step 1) and place it in a 1.5ml centrifuge tube (12000rpm, 10min), pour off the supernatant, and then use sterilized double distilled water After mixing the mycelium, centrifuge (12000rpm, 10min), discard the supernatant, and keep the fungal mycelium;

[0018] 3) Add extraction solution: add 500ul of DNA extraction buffer [2% SDS, 1.4M NaCl...

Embodiment 2

[0025] A simple wall-breaking method for extracting filamentous fungal genome DNA, the method comprising the following steps:

[0026] 1) Filamentous fungus culture: Filamentous fungus B was cultured in NBRIP [National Botanical Research Institute’sphosphatemedium (pH7.0), containing: 10gglucose, 5gCa per liter 3 (PO 4 ) 2 ,5gMgCl 2 ·6H 2 O,0.25gMgSO 4 ·7H 2 O, 0.2gKCl, 0.1g (NH 4 ) 2 SO 4 ] medium, a 250ml Erlenmeyer flask was loaded with 50ml liquid medium, cultured with vibration at 28°C for 2 days, and then cultured statically for 6 days;

[0027] 2) Collect fungal mycelium: Use a sterilized toothpick to pick the fungus B cultured in step 1) and place it in a 1.5ml centrifuge tube (12000rpm, 10min), pour off the supernatant, and then use sterilized double distilled water After mixing the mycelium, centrifuge (12000rpm, 10min), discard the supernatant, and keep the fungal mycelium;

[0028] 3) Add extraction solution: add 500ul of DNA extraction buffer [2% SDS, 1....

Embodiment 3

[0036] A simple wall-breaking method for extracting filamentous fungal genome DNA, the method comprising the following steps:

[0037] 1) Filamentous fungus culture: Cultivate filamentous fungus C in potato medium (20% potatoes, 2% sucrose), fill a 250ml Erlenmeyer flask with 50ml liquid medium, culture at 28°C for 1 day, and then culture it statically for 6 days ;

[0038] 2) Collect fungal mycelium: Pick up the fungus C cultured in step 1) with a sterilized toothpick, put it in a 1.5ml centrifuge tube (12000rpm, 10min), pour off the supernatant, and then use sterilized double distilled water After mixing the mycelium, centrifuge (12000rpm, 10min), discard the supernatant, and keep the fungal mycelium;

[0039] 3) Add extraction solution: add 500ul of DNA extraction buffer [2% SDS, 1.4M NaCl, 0.2MTris-HCl (pH8.0), 0.02MEDTA (pH8.0)], and mix the bacteria and extraction buffer with a sterile toothpick ;

[0040] 4) Freezing: put the bacteria obtained in step 3) into a -20°C...

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

A wall-breaking method for extracting filamentous fungal genome DNA, which uses multi-step centrifugation and glass rod extrusion to break the wall, and specifically cultures the fungus in sequence, collects the bacteria, adds the extract in multiple steps, freezes, and breaks the filamentous bacteria. A method for collecting genomic DNA solution and precipitating genomic DNA. This method uses a -20°C ordinary refrigerator and self-made glass rods, and the experimental equipment is easy to obtain, which reduces the dependence on liquid nitrogen in the traditional method and the contamination of bacteria during mortar grinding. The filamentous fungus genome DNA can be obtained conveniently and quickly, and can be popularized and used.

Description

technical field [0001] The invention relates to a wall-breaking method for genome extraction, in particular to a wall-breaking method necessary for the genome extraction process of filamentous fungi. Background technique [0002] Filamentous fungi widely exist in nature and have a variety of functions. Therefore, the study of filamentous fungi has not declined for a long time, but has attracted more and more attention from researchers. The first step in microbial research is species identification. The traditional classification and identification of fungi is mainly based on the morphological characteristics during culture, and the differences in culture characteristics are very significant, thus affecting the accuracy of classification. The modern taxonomic identification of fungi is a comprehensive identification combining morphology, physiology and DNA molecular level. DNA extraction is the first step in identification at the molecular level, and there are many methods f...

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 陈国华吴广陈汉臣王凤玲
Owner CHINA THREE GORGES UNIV
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