Singh Madhu V, Weil P Anthony
Department of Molecular Physiology and Biophysics, School of Medicine, Vanderbilt University, Nashville, TN 37232-0615, USA.
Anal Biochem. 2002 Aug 1;307(1):13-7. doi: 10.1016/s0003-2697(02)00018-0.
A rapid technique for purifying plasmids from yeast Saccharomyces cerevisiae is described that yields high-quality DNA suitable for bacterial transformation, yeast transformation, and direct DNA sequencing. The method requires only small culture volumes and proprietary bacterial plasmid miniprep kits that allow one to simultaneously prepare a large number of samples in a very short period of time while avoiding the use of toxic organic chemicals. Both yeast single-copy CEN/ARS and high-copy 2micro shuttle plasmids can be isolated using this method. This technique is useful for plasmid purification from yeast two-hybrid experiments as well as yeast genetics and molecular biology experiments.
本文描述了一种从酿酒酵母中快速纯化质粒的技术,该技术可产生适用于细菌转化、酵母转化和直接DNA测序的高质量DNA。该方法仅需少量培养物,并使用专利细菌质粒小量制备试剂盒,可在极短时间内同时制备大量样品,同时避免使用有毒有机化学物质。酵母单拷贝CEN/ARS和高拷贝2μm穿梭质粒均可采用此方法分离。该技术可用于酵母双杂交实验以及酵母遗传学和分子生物学实验中的质粒纯化。