普洱茶发酵过程中微生物总DNA提取方法的比较

Comparison of total microbial DNA extraction methods from Pu-erh tea fermentation samples

为了快速提取普洱茶发酵过程中微生物总DNA,便于分析微生物群落的多样性及演替情况,对比研究了总DNA提取的四种方法-酶法、试剂盒法、超声波法和变性剂法,以16S rRNA区域通用引物(27F和1492R)和18S rRNA区域通用引物(NS1和NS8)对四种方法提取的总DNA分别进行扩增,根据总DNA提取的大小、产量、纯度和PCR扩增产物进行评估。综合各项指标来看,总DNA提取方法以变性剂法为优,其次是试剂盒法、超声波法和酶法。

In order to quickly extract the total microbial DNA,and analyze the diversity and succession of microbial community during Pu-erh tea fermentation,different genomic DNA extraction methods were employed,including enzyme lysis,kit method,ultrasonic lysis and denaturant lysis.PCR amplification was carried out with a bacterial 16S rRNA targeted primer pair（27F and 1492R）and a fungal 18S rRNA targeted primer pair（NS1 and NS8）.The methods of total DNA extraction could be chosen according to size,yield,purity and PCR amplification products.Owing to the four parameters,denaturant lysis was the best,next was kit method,ultrasonic lysis and enzyme lysis.