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Primer sequence for identifying Aspergillus parasiticus and Aspergillus flavus and identification method thereof

A technology of Aspergillus parasiticus and primer sequence, which is applied in the field of molecular biology, can solve laborious and time-consuming problems, and achieve the effect of high speed, high specificity and high reliability

Inactive Publication Date: 2011-03-16
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional identification methods are time-consuming and labor-intensive

Method used

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  • Primer sequence for identifying Aspergillus parasiticus and Aspergillus flavus and identification method thereof
  • Primer sequence for identifying Aspergillus parasiticus and Aspergillus flavus and identification method thereof
  • Primer sequence for identifying Aspergillus parasiticus and Aspergillus flavus and identification method thereof

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Embodiment Construction

[0023] Selected strains

[0024] Alternaria brassicae, A. brassicicola, Fusarium oxysporum, F. graminearum, Rhizotonia cerealis, Grape cinerea cinerea, Penicillium digitatum, Monilinia fructicola and 3 Aspergillus parasitica strains (SD1-1, SX6-6 and ZJHZ27) and 5 Aspergillus flavus strains (SD3-6 , SD5-1, SX6-8, GS10-1 and ZJHZ2).

[0025] The above-mentioned bacterial strains are preserved in the Institute of Biotechnology of Zhejiang University, and can also be separated and purified by conventional plate streaking. The above-mentioned bacterial strains are only used as experimental materials. Of course, other bacterial strains can also be selected, which has no effect on the implementation of the present invention.

[0026] DNA extraction

[0027] Scrape mycelium (100 mg) from the PDA plate with an inoculation needle, place it in a 1.5-mL Eppendorf tube, add 500 μL of DNA extraction lysate (200 mM Tris-HCl, 50 mM EDTA, 20 mM NaCl, 1% SDS, pH8.0), Grind thoroughly with a...

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Abstract

The invention discloses a primer sequence used to identify Aspergillus parasiticus and Aspergillus flavus; the primer sequence comprises two sets of primers; the forward primer in the first set of primers is provided with the base sequence of SEQ ID NO:1 in a sequence table, and the reverse primer of the first set of primers is provided with the base sequence of SEQ ID NO:2 in the sequence table;the forward primer in the second set of primers is provided with the base sequence of SEQ ID NO:3 in the sequence table, and the reverse primer of the second set of primers is provided with the base sequence of SEQ ID NO:4 in the sequence table. The primer sequence has high specificity, identities Aspergillus parasiticus and Aspergillus flavus through PCR amplification with high speed and reliability, thereby laying theoretical foundation for researching the soil polluted by aflatoxin or the distribution and dynamic changes of two types of fungi in crops and providing scientific bases for thecontrol of aflatoxin.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a primer sequence for identifying Aspergillus parasiticus and Aspergillus flavus and an identification method thereof. Background technique [0002] Aspergillus parasiticus and A.flavus have similar phenotypic characteristics and both can produce serious carcinogenic metabolites aflatoxin. Aflatoxins are the most toxic class of biological toxins found so far in contaminated agricultural products. Among them, aflatoxin B1 is the most toxic and harmful, and its toxicity is 10 times that of potassium cyanide and 68 times that of arsenic. It is listed as a highly toxic substance under strict control. Aflatoxin contamination of agricultural products has become an invisible killer affecting food safety and endangering people's health. [0003] At present, it is very difficult to identify Aspergillus parasitica and Aspergillus flavus. Traditional morphological methods are mainly used....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
Inventor 尹燕妮马忠华
Owner ZHEJIANG UNIV
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