Ruppert A, Szalay B, van den Boom D, Horst G, Köster H
Faculty of Chemistry, Department of Biochemistry and Molecular Biology, University of Hamburg, Germany.
Anal Biochem. 1995 Sep 1;230(1):130-4. doi: 10.1006/abio.1995.1447.
A method for the simultaneous isolation of plasmid DNA from as many as 96 Escherichia coli clones in less than 2 h is described. It is based on a modified version of the alkaline lysis procedure originally described by Birnboim and Doly (Nucleic Acids Res. 7, 1513-1523, 1979). The handling of DNA samples is facilitated by the use of microtiter plates with membrane filter bottoms. All centrifugation steps are replaced by filtrations ("the filtration method"). The yield of plasmid DNA from 0.35 ml of an overnight culture is sufficient for restriction analysis of the plasmid clones. Up to 400 nucleotides' readable sequences could be obtained in cycle sequencing reactions with an unmodified sequencing protocol on an automated ABI sequencer.
本文描述了一种在不到2小时的时间内从多达96个大肠杆菌克隆中同时分离质粒DNA的方法。该方法基于Birnboim和Doly最初描述的碱性裂解程序的改进版本（《核酸研究》7，1513 - 1523，1979）。使用带有膜滤器底部的微量滴定板便于处理DNA样品。所有离心步骤均被过滤（“过滤法”）取代。从0.35 ml过夜培养物中获得的质粒DNA产量足以对质粒克隆进行限制性分析。在自动ABI测序仪上使用未修改的测序方案进行循环测序反应时，可获得多达400个核苷酸的可读序列。
