| PCR法检测耐热耐酸菌条件的优化 |
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| 引用本文： | 王宏,常玉华,仇农学. PCR法检测耐热耐酸菌条件的优化[J]. 食品与发酵工业, 2004, 30(11): 99-101,105 |
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| 作者姓名： | 王宏 常玉华 仇农学 |
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| 作者单位： | 陕西师范大学食品工程系,西安,710062 |
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| 基金项目： | 国家科技部“十五”星火计划资助项目 (2 0 0 1EA85 0 0 30 ) |
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| 摘 要： | 以耐热菌为研究对象 ,对PCR法快速检测耐热菌的扩增条件 :Taq酶浓度、Mg2 +浓度、退火温度、循环温度 /时间、循环次数 5个方面进行了优化。最终得到PCR反应最佳条件 :5 0 μL扩增体系中 ,Taq酶浓度 2U/mL、Mg2 +浓度 2 0mmol/L、退火温度 5 8℃。扩增程序 :94℃预变性 4min后进入PCR循环 ,即 94℃ /3 0S -5 8℃ /3 0S -72℃ /3 0S ,循环 3 5次 ,72℃下延伸 5min。
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| 关 键 词： | PCR 耐热菌 条件优化 |
Optimization of PCR Reaction Conditions in Detection of Alicyclobacillus acidoterrestris |
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| Wang Hong Chang Yuhua Qiu Nongxue. Optimization of PCR Reaction Conditions in Detection of Alicyclobacillus acidoterrestris[J]. Food and Fermentation Industries, 2004, 30(11): 99-101,105 |
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| Authors: | Wang Hong Chang Yuhua Qiu Nongxue |
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| Abstract: | Study on the reaction conditions i. e. concentration of Taq DNA polymerase and Mg 2+ , annealing temperature, recycle temperature/time, number of cycles in rapid detection of Alicyclobacillus acidoterrestris by PCR technology was presented in this paper. The optimized parameters achieved for 50μL PCR reaction system are 2?U/mL Taq DNA polymerase , 2.0?mmol/L [Mg 2+ ], 58℃ annealing tempreture. The amplification sequences is: pre-denaturation at 94℃ for 4?min, then run the PCR for 35 cycles including 94℃ for 30s～58℃ for 30s ～72℃ for 30s, and finally extend the process at 72℃ for 5?min. |
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| Keywords: | PCR Alicyclobacillus acidoterrestris optimizing reation conditions |
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