| 研究土壤微生物多样性的PCR条件优化 |
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| 引用本文： | 邸宁,刘志民,马焕普.研究土壤微生物多样性的PCR条件优化[J].安徽农业科学,2011,39(23):14059-14061,14064. |
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| 作者姓名： | 邸宁 刘志民 马焕普 |
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| 作者单位： | 北京农学院,北京,102206 |
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| 基金项目： | 北京市教委资助项目(KM200910020001) |
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| 摘 要： | 目的]对PCR-DGGE法的试验条件进行优化,以更好地分析土壤微生物遗传多样性。方法]改良高盐法提取土壤DNA,改进引物设计,改变PCR反应过程中退火温度、扩增体系,比较PCR扩增结果。结果]经过改良的高盐法提取的土壤微生物DNA效果更好。PCR扩增选择20μl体系条带单一,易于操作。退火温度选择在55℃时无非特异性扩增,35个循环次数易于后续DGGE分析。结论]已经优化的PCR基因扩增体系特异性高且稳定可靠。
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| 关 键 词： | PCR-DGGE 土壤微生物 多样性 |
Optimization of PCR Condition in Soil Microbial Diversity Study |
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| Institution: | DI Ning et al(Agriculture University of Beijing,Beijing 102206) |
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| Abstract: | Objective] The research aimed to optimize the condition of PCR to analyze genetic diversity of soil microorganism.Method] Different PCR systems were set up by altering the factors,such as primer design,volume,annealing temperature and cycles.Result]In the optimized PCR system,specific,sensitive and stable amplified bands were obtained when cycles were 35,the annealing temperature was 55 ℃,the volume of PCR was 20 μl.Conclusion] The optimized PCR reaction system had high specificity and reliability. |
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| Keywords: | PCR-DGGE Soil microorganisms Diversity |
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