摘要
本文报导了一种较为简单的入噬菌体DNA制备方法.首先在LB琼脂平皿中扩增噬菌体,待取得1×10^(11)噬菌体后进行氯化铯梯度离心,取出噬菌体条带;继而加入蛋白酶K分解噬菌休颗粒以释放其DNA;经酚/氯仿抽提和酒精沉淀,最后可得纯DNA10-20ug。该样品可用于限制酶酶谱分析或次级克隆。
An improved methos for λ bacteriophage DNA prepartion was reported. The DNA was prepared from a phage plate lysate after amplifying the phage to 1×10^(11) pfu, a CsCl step gradient centrifugation was carried out at 35000rpm for two hours. Then the phage band was recovered by a Syringe with a 25-G needle,and lysis of phage particles with proteinase K. were performed Phenol/chloroform extraction and ethanol precipitation were used for further purification. A yield of 10-20ug of the phage DNA was obtained from 3 plates. The preparaed DNA is pure enough for restriction analysis and subcloning.
出处
《南京大学学报（自然科学版）》
CAS
CSCD
1991年第3期515-519,共5页
Journal of Nanjing University（Natural Science）
关键词
Λ噬菌体
脱氧核糖核酸
克隆
bacteriophage λDNA
gradient centrifugation
cloning
