[Rapid isolation of phage lambda DNA].
A modified procedure in two versions (micro, for 10 ml of phage lysate, and macro, for 200-500 ml) is described for preparing lambda phage DNA. The main advantage of the modified method is that it gives a possibility to isolate high-quality DNA from lambda phage lysates in 2-3 hrs. Only standard solutions (TE, NaCl, SDS, MgCl2, EDTA, RNAse A) were used throughout the whole protocol. Incubation with DNAse I and proteinase K was omitted and in microvariant concentration of the phage by PEG 6000 was excluded. Digestion by RNAse A was performed in solution with EDTA and SDS and leads to RNA degradation. The yields of DNA (0.5-2 micrograms per ml of L-broth) are similar to those obtained by other methods. DNA quality is better than in the samples of DNA prepared by other express-methods and practically the same as after CsCl centrifugation. DNA can be used for splitting by restriction enzymes, cloning and gene library construction.
Zabarovskiĭ
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Turina
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D010582:Bacteriophage lambda / Q000235:genetics*; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000302:isolation & purification*; D004492:Edetic Acid; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics; D012259:Ribonuclease, Pancreatic; D012967:Sodium Dodecyl Sulfate
D004279:DNA, Viral; D012967:Sodium Dodecyl Sulfate; D004492:Edetic Acid; D004262:DNA Restriction Enzymes; D012259:Ribonuclease, Pancreatic
Bystroe vydelenie DNK faga lambda.