Gibson T J, Sulston J E
MRC Laboratory of Molecular Biology, Cambridge, England.
Gene Anal Tech. 1987 May-Jun;4(3):41-4. doi: 10.1016/0735-0651(87)90016-1.
A protocol is described for the growth and preparation of plasmid DNAs from small culture volumes (250 microliters) and utilizing standard 96-well plates. Several hundred plasmids can be prepared simultaneously, yielding sufficient DNA for subsequent analysis by restriction digestion and gel electrophoresis. This protocol may be useful for rapid screening of clones arising in recombinant DNA work such as site-directed mutagenesis, oligonucleotide cassette cloning, deletion analysis, etc. The technique was initially developed to meet our requirement to provide large numbers of cosmid DNAs for restriction enzyme fingerprint analyses in genome mapping projects.
本文描述了一种从少量培养体积（250微升）中培养和制备质粒DNA的方法，该方法使用标准的96孔板。可同时制备数百个质粒，产生足够的DNA用于后续的限制性酶切消化和凝胶电泳分析。该方法对于快速筛选重组DNA工作中产生的克隆（如定点诱变、寡核苷酸盒式克隆、缺失分析等）可能有用。该技术最初是为满足我们在基因组图谱绘制项目中提供大量黏粒DNA用于限制性酶切指纹分析的需求而开发的。
