A procedure for large-scale isolation of RNA-free plasmid and phage DNA without the use of RNase.
A preparative procedure for the large-scale isolation of plasmid DNA without the use of RNAse is described. Crude plasmid DNA is prepared using a standard boiling method. High-molecular-weight RNA is removed by precipitation with LiCl, and low-molecular-weight RNA is removed by sedimentation through high-salt solution. The procedure is inexpensive, rapid, simple, and particularly suitable for processing several large-scale preparations simultaneously. A similar procedure has been developed for preparation of lambda-phage DNA.
D010582:Bacteriophage lambda / Q000032:analysis*; D002499:Centrifugation, Density Gradient; D011232:Chemical Precipitation; D002712:Chlorides; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004279:DNA, Viral / Q000302:isolation & purification*; D004926:Escherichia coli / Q000032:analysis / Q000235:genetics; D008094:Lithium; D018021:Lithium Chloride; D010957:Plasmids; D012329:RNA, Bacterial / Q000302:isolation & purification; D012367:RNA, Viral / Q000302:isolation & purification; D012260:Ribonucleases
D002712:Chlorides; D004269:DNA, Bacterial; D004279:DNA, Viral; D012329:RNA, Bacterial; D012367:RNA, Viral; D008094:Lithium; D012260:Ribonucleases; D018021:Lithium Chloride