A rapid method for the preparation of plasmid DNA.
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.
Srivastava
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Department of Pediatrics, Washington University School of Medicine, Saint Louis, MO 63110, USA.
D004247:DNA / Q000302:isolation & purification*; D010957:Plasmids / Q000235:genetics*; D013997:Time Factors
D004247:DNA