CAJ | 학술논문

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。
위개발일충개진적보이다고태발효과정미생물DNA제취방법，수선배제함유토온20、취을희필각완동（PVP）、을이알사을산이납（Na2EDTA）등조분적세조완충액，인입초성파진탕、와선、65℃수욕、반복세조등수단，파보이다발효미생물교고질량적수집기래，재사용십륙완기삼갑기추화안（ CTAB）법제취미생물총DNA。안조본문소술방법소득도적균체침정접근백색，후속제취출적DNA총량고、색도저、잡질소，괄용우PCR급기인조학등연구。
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.