A rapid and simple method for screening large numbers of recombinant DNA clones.
A simple and rapid method has been described for the isolation of plasmid, phagemid and phage DNAs. Hundreds of recombinant clones can be screened in one day employing this method. It takes half an hour to prepare plasmid DNA from ten clones, and the DNA prepared from a single colony using this method is of sufficient quality and in sufficient amount to perform at least five restriction digestions. This method eliminates the need for RNase treatment and phenol chloroform extraction if the plasmids are needed only for the restriction digestion. If needed, RNAs can be removed after restriction digestion by adding RNase and incubating for two minutes at room temperature. After RNase treatment and phenol/chloroform extraction, the plasmid DNA serves as a good template for sequencing. The DNA can be stored at -20 degrees C for over eight weeks.
Srivastava
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RA
Dept. of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63110.
Schonfeld
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G
D000818:Animals; D001055:Apolipoproteins B / Q000235:genetics; D001435:Bacteriophages / Q000032:analysis; D002725:Chloroform; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics; D019800:Phenol; D010636:Phenols; D010957:Plasmids; D051381:Rats; D012260:Ribonucleases; D013698:Templates, Genetic; D014169:Transformation, Bacterial
D001055:Apolipoproteins B; D004274:DNA, Recombinant; D010636:Phenols; D019800:Phenol; D002725:Chloroform; D004247:DNA; D012260:Ribonucleases