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用十二烷基硫酸钠碱裂解法制备质粒DNA:小量制备

Preparation of Plasmid DNA by Alkaline Lysis with Sodium Dodecyl Sulfate: Minipreps.

作者信息

Green Michael R, Sambrook Joseph

出版信息

Cold Spring Harb Protoc. 2016 Oct 3;2016(10):2016/10/pdb.prot093344. doi: 10.1101/pdb.prot093344.

Abstract

In this protocol, plasmid DNA is isolated from small-scale (1-2 mL) bacterial cultures. Yields vary between 100 and 5 µg of DNA, depending on the copy number of the plasmid. Miniprep DNA is sufficiently pure for use as a substrate or template in many in vitro enzymatic reactions. However, further purification is required if the plasmid DNA is used as the substrate in sequencing reactions.

摘要

在本实验方案中,从小规模(1 - 2 mL)细菌培养物中分离质粒DNA。产量在100至5 μg DNA之间,具体取决于质粒的拷贝数。小量制备的DNA纯度足以用作许多体外酶促反应的底物或模板。然而,如果将质粒DNA用作测序反应的底物,则需要进一步纯化。

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