| 窖泥微生物群落SSCP分析条件优化的研究 |
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| 引用本文： | 卫春会,黄治国,甄攀.窖泥微生物群落SSCP分析条件优化的研究[J].四川理工学院学报(自然科学版),2010,23(6). |
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| 作者姓名： | 卫春会 黄治国 甄攀 |
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| 基金项目： | 四川省应用技术研究与开发项目，自贡市2008年重点科技计划项目，酿酒生物技术及应用四川省重点实验室项目 |
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| 摘 要： | 由于窖泥微生物群落结构复杂,而PCR-SSCP是研究环境微生物群落较理想的一种方法。试验对影响SSCP图谱的条件进行优化分析,为窖泥微生物群落结构解析提供了有效的手段。结果表明:变性缓冲液配比为:去离子甲酰胺1.9mL、溴酚蓝3mg、5×TBE 250μL、0.5M EDTA5μL、10%SDS5μL;凝胶条件为:凝胶浓度为12%、丙烯酰胺与N,N′-亚甲基双丙烯酰胺的比例为29∶1、无甘油。经95℃变性10min,4℃条件下200V电泳8h,可得到较理想的图谱。
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| 关 键 词： | 窖泥 PCR-SSCP 16S rDNA 微生物群落 |
Study on Optimal Conditions of SSCP Analysis Technology for the Microbial Community in Pit Mud |
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| WEI Chun-hui,HUANG Zhi-guo,ZHEN Pan.Study on Optimal Conditions of SSCP Analysis Technology for the Microbial Community in Pit Mud[J].Journal of Sichuan University of Science & Engineering:Natural Science Editton,2010,23(6). |
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| Authors: | WEI Chun-hui HUANG Zhi-guo ZHEN Pan |
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| Abstract: | The microbial community structure of pit mud is complex,but PCR-SSCP is a more ideal method on environmental microbial community.The experiment analyzed conditions of SSCP profile,providing effective means for analysising microbial community structure.The results indicates that: Loading buffer was deionzed formamide being 1.9mL,bromophenol blue being 3mg,5×TBE 250μL,0.5M EDTA 5μL and 10% SDS 5μL;Condition of gel was concentration being 12%,the proportion of acrylamide and N,N'-methylene-bis-acrylamide being 29∶1,without glycerine.Denatured at 95℃ for 10min,4℃,200V,electrophoresis 8h,finally get perfect spectrogram. |
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| Keywords: | pit mud PCR-SSCP 16S rDNA microbial community |
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