一种改进的普洱茶固态发酵过程微生物DNA提取方法

Modified extraction total microbial DNA in Puer tea solid-state fermention

为开发一种改进的普洱茶固态发酵过程微生物DNA提取方法,首先配制含有吐温-20、聚乙烯吡咯烷酮(PVP)、乙二胺四乙酸二钠(Na2EDTA)等组分的洗涤缓冲液,引入超声波振荡、涡旋、65℃水浴、反复洗涤等手段,把普洱茶发酵微生物较高质量的收集起来,再使用十六烷基三甲基溴化铵(CTAB)法提取微生物总DNA。按照本文所述方法所得到的菌体沉淀接近白色,后续提取出的DNA总量高、色度低、杂质少,适用于PCR及基因组学等研究。

Isolation high purity, large fragment DNA is important in microbial molecular research and while collecting microbial cells properly plays key roles to isolate total DNA？We developed a modified method to extract total microbial DNA in Puer tea fermentation？First, microbes tea was collected by washing samples several times with a buffer containing with Tween？20,polyvinylpyrrolidone（PVP）,and ethylenediaminetetraacetic acid disodium （ Na2 EDTA ） , in combination with ultrasonic oscillations and vortex at 65 ℃ until microbial cells with nearly white color. Then,microbial total DNA was extracted by using hexadecyltrimethylammouium bromide（ CTAB） approach to obtain an integral,light color and pure microbial DNA suitable for PCR and genomics research.