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Position: Home > Articles > Study on DNA Extraction and PCR-RFLP Detection Method of Aspergillus flavus FOOD SCIENCE 2008,29 (12) 393-396

黄曲霉DNA提取及PCR-RFLP检测方法的建立

作  者:
邵碧英;王传得;郑晶;黄晓蓉
单  位:
福建农林大学动物科学学院;福建出入境检验检疫局
关键词:
黄曲霉;DNA;PCR-RFLP
摘  要:
采用3种提取方法、3种菌丝前处理提取黄曲霉标准菌株的DNA,凝胶电泳结果表明经-80℃冷冻过夜或液氮处理菌丝、用氯化苄方法提取、并经RNase处理的DNA效果最好。合成扩增aflR基因的2对引物,用巢式PCR验证PCR产物的非假阳性。PCR产物的HincⅡ和PvuⅡ酶切结果与预期的完全一致,表明建立的黄曲霉PCR-RFLP检测方法是可行的。
译  名:
Study on DNA Extraction and PCR-RFLP Detection Method of Aspergillus flavus
作  者:
SHAO Bi-ying1,WANG Chuan-de2,ZHEN Jing1,HUANG Xiao-rong1(1. Fujian Entry-Exit Inspection and Quarantine Bureau,Fuzhou 350001,China; 2. College of Animal Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
关键词:
Aspergillus flavus;DNA;PCR-RFLP
摘  要:
Aspergillus flavus standard strain DNAS were isolated by using three extraction methods and three pretreatment methods of mycelia for benzyl chloride extraction method. The gel electrophoresis results showed that the DNA isolated by using benzyl chloeide method and treated with RNase is the best after the mycelia are freezed overnight at -80 ℃ or grinded with liquid nitrogen. Two pairs of primes were synthesized to amplify aflR gene. The PCR products are not false positive which is confirmed by nest-PCR. The expected restriction endonuclease analysis results of the PCR products with HincⅡand PvuⅡwere achieved,which showed that the PCR-RFLP detection method is feasible for Aspergillus flavus.

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