【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。更多还原

【Abstract】 [Objective] The research aimed to optimize the test condition of PCR-DGGE method to analyze genetic diversity of soil microorganism.[Method] The amplified results of PCR were compared by improving high salt method for extracting soil DNA,improving primer design,changing the annealing temperature and amplification system of PCR reaction process.[Result] The result of extracting soil microbial DNA was better by high salt method which was improved.A single band was obtained and operation was easy when choosing 20 μl system for PCR amplification.There was no nonspecific amplification when choosing annealing temperature at 55 ℃,and cycle number of 35 was easy for following DGGE analysis.[Conclusion] The optimized PCR reaction system has high specificity and reliability.更多还原