Internal reference gene of cordyceps militaris mycelium under cold stress, and primers, screening method and application of internal reference gene

A technology of Cordyceps militaris mycelium and internal reference genes, which is applied in biochemical equipment and methods, microbe measurement/inspection, DNA/RNA fragments, etc. It can solve the problems of no internal reference gene research disclosure and achieve high efficiency of primer amplification , low cost and simple screening method
CN110791585AActive Publication Date: 2020-02-14CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
Publication Date
2020-02-14

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Abstract

The invention discloses an internal reference gene of the cordyceps militaris mycelium under cold stress, and primers, screening method and application of the internal reference gene. The internal reference gene is UBC, a nucleotide sequence is SEQ ID NO: 1, nucleotide sequences of the forward and reverse primers of the UBC are SEQ ID NO: 13-14. The screening method is as follows: (1) selecting 12internal reference genes as candidate internal reference genes, and designing primers; (2) treating cordyceps militaris under cold stress, extracting the total RNA from the mycelium, measuring the concentration of RNA, synthesizing cDNA through reverse transcription of RNA, using cDNA as a template, and analyzing the candidate internal reference genes by using real-time fluorescence quantitativePCR; and (3) performing Ct value analysis on the real-time fluorescence quantitative PCR data to screen the stable internal reference gene. The application is that the UBC is used as the internal reference gene, and the relative expression of the target gene under cold stress is detected by using real-time fluorescence quantitative PCR. The internal reference gene has good stability. The method issimple, fast, and low in cost.
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Description

technical field

[0001] The present invention relates to an internal reference gene, primers, screening method and application, in particular to an internal reference gene, primer, screening method and application of Cordyceps militaris mycelia under cold stress. Background technique

[0002] Cordyceps militaris is a model species of Ascomycota, Hypocreta, Ergotaceae, and Cordyceps. Its scientific name is Cordycepsmilitaris. The fruiting bodies of Cordyceps militaris have anti-inflammatory, antibacterial, anti-tumor, anti-aging and immunomodulatory effects. At present, using insects (such as silkworm chrysalis), rice, etc. as the main raw materials of the medium, a commercial production method for planting fruiting bodies of Cordyceps militaris has been established. However, little research has been done on the molecular mechanisms underlying pupal fruiting body formation, which is a fundamental biological question.

[0003] Temperature is one of the key factors affecting t...

Claims

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