Volvariella volvacea strain 0229 molecule specific detection markers and detection method using the same

A technology for detecting markers and detection methods, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. , the detection time is short, the effect of speeding up the research and development work

Active Publication Date: 2014-06-04
SHANGHAI ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Our country has a wide area of ​​straw mushroom cultivation, and its output once ranked fifth among edible fungi, but the variety is single. After testing the mating gene types of the main cultivated straw mushroom strains, it was f

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  • Volvariella volvacea strain 0229 molecule specific detection markers and detection method using the same
  • Volvariella volvacea strain 0229 molecule specific detection markers and detection method using the same

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Embodiment 1

[0023] DNA extraction:

[0024] Inoculate the PDA plate medium with the strain of straw mushroom, culture it at 32°C for 5 days, scrape the surface mycelia and put it into a sterile centrifuge tube, store it in a -70°C refrigerator, and then use the improved CTAB method to extract genomic DNA (the method is briefly described as follows: Grind the freeze-dried mycelia into powder, add 2×CTAB extract preheated at 65°C, dispense into 2mL centrifuge tubes, keep warm at 65°C for 45min, and shake gently occasionally; centrifuge at 4°C, 12000rpm for 20min, and take the supernatant Add an equal volume of chloroform: isoamyl alcohol (24:1) mixture, mix gently, and centrifuge at 12000rpm for 20min at 4°C after 15min. Take the supernatant and transfer it to a new centrifuge tube, add 2 / 3 volume -20°C Pre-cooled isopropanol, shake gently, centrifuge at 4°C, 8000rpm for 10min after 5min. Discard the supernatant, wash the pellet with 75% ethanol (containing 10mmol / L NaAc) three times, centr...

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Abstract

The invention relates to volvariella volvacea strain 0229 molecule specific detection markers and a detection method using the same. The volvariella volvacea strain 0229 molecule specific detection markers are specific primers and sequences of the specific primers comprise F: CTTTCAGGAAGAGCCCAGC and R: TGAAGGGCTTGAAGGAGAAA. The detection method comprises the following steps of inoculating a PDA flat medium with a volvariella volvacea strain, carrying out culture at a temperature of 32 DEG C for 5 days, collecting surface hyphae by scraping, putting the surface hyphae into a sterile centrifuge tube, carrying out preservation at a temperature of -70 DEG C in a refrigerator, extracting a genome DNA by an improved CTAB method to obtain a DNA, carrying out PCR amplification by the DNA as a template and specific detection marker primers as amplimers to obtain a PCR amplification product, and carrying out electrophoresis detection. The volvariella volvacea strain 0229 molecule specific detection markers have the advantages of short detection time and high accuracy, are used for molecular marker-based identification of volvariella volvacea strain 0229, are conducive to volvariella volvacea breeding development and accelerate research and development of a novel volvariella volvacea variety.

Description

technical field [0001] The invention belongs to the field of detection marks and detection methods of straw mushroom strains, and in particular relates to a molecular-specific detection mark of straw mushroom strain 0229 and a detection method thereof. Background technique [0002] Straw mushroom (Volvariella volvacea) cultivation originated in China, has a cultivation history of more than 300 years so far, and is widely cultivated in southern my country. Straw mushrooms are delicious, not only rich in nutritional value, but also have an important role in health care, and are deeply loved by consumers. Straw mushroom is a high-temperature cultivated edible fungus. Compared with other edible fungi, its remarkable feature is that its growth rate is extremely fast. It usually only takes 2 weeks from sowing to harvesting. Its annual output has always ranked first in my country's edible fungi industry. Top ten. [0003] Our country has a wide area of ​​straw mushroom cultivation...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 汪虹王莹陈明杰鲍大鹏冯爱萍
Owner SHANGHAI ACAD OF AGRI SCI
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