溶解
苯酚萃取
λ噬菌体
DNA
琼脂糖
噬菌体
重组DNA
分子生物学
限制性酶
色谱法
化学
DNA提取
噬菌体
生物化学
生物
大肠杆菌
聚合酶链反应
基因
核糖核酸
作者
Jeremy W. Dale,Peter Greenaway
出处
期刊:Humana Press eBooks
[Humana Press]
日期:2003-11-14
卷期号:: 211-216
被引量:3
标识
DOI:10.1385/0-89603-064-4:211
摘要
The isolation of phage DNA from a purified phage preparation simply involves the removal of the proteins of the phage particle. This is done most easily by phenol extraction (Method A). The large amounts of high quality DNA needed for use as vectors or for markers for gel electrophoresis can be obtained in this way. However, sometimes (for example, when screening recombinant phages) only small amounts of phage DNA are required. The purity of these samples needs only to be sufficient for the restriction enzyme digests to provide distinct patterns on an agarose gel. Method B provides one way in which this can be done (1). A plate lysate is prepared and the released phage is subsequently lysed by the addition of SDS. The phage proteins and SDS are precipitated by the addition of potassium acetate to leave the phage DNA in solution. Finally, the DNA is recovered by ethanol precipitation.
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