Srivastava N
Department of Pediatrics, Washington University School of Medicine, Saint Louis, MO 63110, USA.
Indian J Exp Biol. 1995 May;33(5):387-91.
A rapid method was developed to prepare plasmid DNA to be used for in vitro transcription, subcloning and sequencing. Clean plasmid DNA can be prepared within 90 min using this method and the quality of the plasmid DNA was found to be suitable for sequencing, preparation of in vitro RNA transcript and electroporation. Using this method plasmid DNA can be prepared from 2 ml to 100 ml culture. The yield of the recombinant plasmids was found to be in the range of 50-100 micrograms from 10 ml culture depending on the low and high copy number plasmids. The advantage of this method is that one can easily and rapidly prepare large quantities of plasmid DNA of high quality to be used for various purposes.
开发了一种快速方法来制备用于体外转录、亚克隆和测序的质粒DNA。使用该方法可在90分钟内制备出纯净的质粒DNA，且发现该质粒DNA的质量适用于测序、体外RNA转录制备和电穿孔。使用此方法可从2毫升至100毫升培养物中制备质粒DNA。根据低拷贝数和高拷贝数质粒的不同，从10毫升培养物中获得的重组质粒产量在50至100微克范围内。该方法的优点是可以轻松快速地制备大量高质量的质粒DNA，用于各种目的。
