Pickard Derek John Juan
The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.
Methods Mol Biol. 2009;502:3-9. doi: 10.1007/978-1-60327-565-1_1.
Preparation of pure bacteriophage DNA used to rely on using CsCl gradients to give high purity or methods that yielded DNA that was either of low recovery or subject to significant genomic contamination. Recently though, new methods have come along that allow the purification of DNA from plate lysates that are not only capable of high yield but also, for all intents and purposes, free of genomic contamination (i.e. no visible genomic contamination on restriction analysis or when used for bacteriophage sequencing). This protocol that form the basis of this short section can be used to prepare bacteriophage DNA from one or two 9 cm L-agar plates. For these preps, the use of agarose in the top agar is recommended to avoid any restriction inhibitors that may be present in some agar preparations.
过去，制备纯噬菌体DNA依赖于使用氯化铯梯度以获得高纯度，或者采用回收率低或易受显著基因组污染的方法。然而，最近出现了新的方法，能够从平板裂解物中纯化DNA，不仅产量高，而且在所有实际应用中均无基因组污染（即限制性分析时或用于噬菌体测序时无可见的基因组污染）。构成本简短章节基础的此方案可用于从一两个9厘米的L-琼脂平板制备噬菌体DNA。对于这些制备，建议在顶层琼脂中使用琼脂糖，以避免某些琼脂制剂中可能存在的任何限制性抑制剂。
