Green Michael R, Sambrook Joseph
Cold Spring Harb Protoc. 2017 Nov 1;2017(11):pdb.prot093443. doi: 10.1101/pdb.prot093443.
The double-stranded, closed-circular, replicative form (RF) of M13 DNA is present in high copy numbers in infected cells, and its physical characteristics are essentially identical to those of closed-circular plasmid DNAs. Any of the methods commonly used to purify plasmid DNA can therefore be used to isolate M13 RF DNA. This protocol describes the isolation of M13 RF DNA by alkaline lysis from small volumes (1-2 mL) of infected bacterial cultures. The yield of DNA (1-4 mg, depending on the size of the M13 clone) is more than enough for most purposes in molecular cloning. However, should more DNA be needed, the procedure can easily be scaled up.
M13 DNA的双链闭环复制型（RF）在感染细胞中以高拷贝数存在，其物理特性与闭环质粒DNA基本相同。因此，任何常用于纯化质粒DNA的方法均可用于分离M13 RF DNA。本方案描述了通过碱裂解法从少量（1 - 2 mL）感染细菌培养物中分离M13 RF DNA的方法。DNA产量（1 - 4 mg，取决于M13克隆的大小）对于分子克隆中的大多数用途来说绰绰有余。然而，如果需要更多DNA，该程序可以很容易地扩大规模。
