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。
[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.
出处
《安徽农业科学》
CAS
北大核心
2011年第23期14059-14061,14064,共4页
Journal of Anhui Agricultural Sciences
基金
北京市教委资助项目(KM200910020001)
