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

Method for extracting edible mushroom DNA

A technology for edible fungi and mycelium, applied in the field of edible fungi genetic engineering, can solve the problems of time-consuming and expensive, long extraction time, and high extraction cost, and achieve the effects of reducing test costs, good DNA integrity, and no environmental pollution.

Inactive Publication Date: 2014-03-12
SHANGHAI ACAD OF AGRI SCI
View PDF2 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The entire extraction process is cumbersome, time-consuming and costly. Although there are some simplified methods, they either increase the cost or require the assistance of other test tools.
In short, the current edible fungus DNA extraction methods generally have cumbersome steps, long extraction time, and high extraction costs. There is no DNA preparation method that can be completed with only a simple reagent and simple operation.

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
  • Method for extracting edible mushroom DNA
  • Method for extracting edible mushroom DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1. Preparation of DNA from mushrooms, oyster mushrooms, enoki mushrooms, Agaricus bisporus, black fungus and Grifola frondosa.

[0026] 1. Materials

[0027] Lentinula edodes, Pleurotus ostreatus, Flammulina velutiper, Agaricus bisporus, Auricularia auricula-judae and Griflola frondosa varieties were all from Shanghai Academy of Agricultural Sciences The culture preservation center of the Institute of Edible Mushrooms preserves the strains, and the preservation numbers are 4627, 0860, 0188, 0897, 0115 and 2611.

[0028] 2. Reagents

[0029] TE buffer: 10mmol / L Tris-HCl, 1mmol / L EDTA, pH8.0.

[0030] 3. DNA extraction

[0031] (1) Materials: Lentinus edodes, oyster mushroom, Flammulina velutipes, Agaricus bisporus, black fungus and Grifola frondosa mycelia were cultured and grown on PDA plates for 2 days, and the young aerial mycelia were scraped on the surface of the plate with a sterilized toothpick for 2~ 3 times until there is a small group of hyphae v...

Embodiment 2

[0034] Embodiment two, DNA content and purity detection

[0035] The DNA extracted in Example 1 was detected by a Thermo Company NanoDrop 1000 spectrophotometer for nucleic acid content and purity, and 0.1 g of mushroom hyphae DNA extracted by the CTAB method was used as a control. The test results show:

[0036] (1) The nucleic acid concentrations of shiitake mushroom, oyster mushroom, Flammulina velutipes, Agaricus bisporus, black fungus, and Grifola frondosa were 65, 114, 140, 100, 53, and 27 ng / μL, respectively, with Grifola frondosa being the lowest and Flammulina velutipes the highest. The mushroom DNA extracted by the CTAB method is 730ng / μL, which needs to be diluted for conventional PCR reactions;

[0037] (2) The results of OD260 / 280 showed that the ratio of OD260 / 280 of shiitake and oyster mushrooms was between 2.0 and 2.2, which was comparable to that of the control, and that of the other four species were all between 1.6 and 2.0, indicating that there were protei...

Embodiment 3

[0039] Embodiment three, multiple PCR reaction verification

[0040] The mushroom DNA extracted in Example 1 was directly subjected to multiple PCR reactions without dilution to verify whether the content and purity of the DNA samples would meet the requirements of common PCR reactions. The DNA extracted by the CTAB method was diluted to a concentration of 50 ng / μL as a control, the ITS fragment was selected as a specific fragment amplification test, and RAPD, SSR, ISSR and SRAP were used for various molecular marker amplification tests. The primer sequences used are shown in Table 1. The PCR products were electrophoresed by PAGE and stained by silver staining. For the amplification effects of various PCR reactions, see figure 1 .

[0041] Table 1: ITS, RAPD, SSR, ISSR and SRAP primer sequences for PCR validation

[0042]

[0043] The PCR amplification effect shows that compared with the DNA extracted by the traditional CTAB method, the mushroom DNA extracted in Example 1...

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 discloses a method for extracting edible mushroom DNA. The method comprises the following steps: (1) getting material: putting edible mushroom hyphae into a buffer solution; (2) heating: heating at the temperature of 90 to 110 DEG C; (3) centrifuging: centrifuging at the speed of 7000 to 9000 rpm for 5 to 10 min, and then extracting a supernatant liquor, wherein the supernatant liquor contains total DNA. The method for extracting the edible mushroom DNA is short in used time, simple and convenient to operate, low in cost and free from contamination to the environment, the prepared DNA is good in integrality and suitable for PCR reaction.

Description

technical field [0001] The invention relates to the field of genetic engineering of edible fungi, in particular to a method for extracting edible fungus DNA. Background technique [0002] In recent years, my country's edible fungus industry has developed rapidly, and the basic research on edible fungi has also penetrated to the molecular level. In the research of classification, genetics, breeding, strain identification, genetic diversity, etc. of edible fungi, DNA needs to be extracted or prepared. These DNAs are used in genome sequencing, PCR amplification of a certain gene or fragment, application of DNA molecular markers, and genetic engineering research. A large proportion of DNA prepared in these researches is used in various PCR reactions. [0003] At present, there are differences in the methods of extracting DNA from various edible fungi mainly in terms of wall breaking, impurity removal and nucleic acid precipitation. The usual method of breaking the wall includes ...

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): C12N15/10
Inventor 章炉军尚晓冬巫萍谭琦宋春艳张丹于海龙
Owner SHANGHAI ACAD OF AGRI SCI
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