发酵肉制品总微生物RAPD扩增条件的优化  

Optimization of RAPD conditions for total microbes in fermented meat

作　　者：秦丹[1] 杨勇[1] 周小平[1] 程燕[1] 

机构地区：[1]四川农业大学食品学院,四川雅安625014

出　　处：《中国酿造》2011年第6期92-96,共5页China Brewing

基　　金：四川省教育厅项目(07ZS004)

摘　　要：以改进的氯化苄法抽提发酵肉制品总微生物DNA,进行随机扩增多态DNA(random amplified polymorphic DNA,RAPD)分析。采用单因子梯度试验法对影响发酵肉制品微生物RAPD反应的反应体系、Mg2+浓度、dNTPs浓度、引物浓度、Taq DNA聚合酶的用量及DNA模板浓度进行了筛选。结果表明,发酵肉制品微生物RAPD扩增条件为25μL PCR反应体积中,2.5mmol/L MgCl2,0.15mmol/LdNTPs,15pmol引物,50ng模板DNA,1.0U Taq DNA聚合酶。The total microbial DNA was extracted by the improved benzyl chloride method from fermented meat and was analyzed by random amplified polymorphic DNA （RAPD） markers. The influence of the concentration of Mg2＋, dNTPs, primer, Taq DNA polymerase and the templates on RAPD-PCR reaction system were investigated by single-factor gradient test method. The optimal conditions were as follows： MgC12 concentration 2.5mmol/L, dNTPs concentration 0.15mmol/L, primer concentration 15pmol, DNA template 50ng and Taq DNA polymerase 1.0U in 25μL RAPD- PCR reaction system.

关 键 词：发酵肉制品 总微生物 随机扩增多态DNA PCR反应体系优化 

分 类 号：TS201.3[轻工技术与工程—食品科学]