Filter-supported preparation of lambda phage DNA.
A rapid and simple method is described for the isolation of DNA from phage lambda which requires neither special equipment nor expensive material such as cesium chloride for ultracentrifugation nor extractions with organic solvents or ethanol precipitation. Microgram quantities of lambda DNA are obtained in less than 2 h from 90-mm plate lysates or 5-ml liquid cultures. The method allows the simultaneous isolation of large numbers of probes, e.g., clones from phage libraries. Lambda phages are precipitated by polyethylene glycol/sodium chloride and recovered by low speed centrifugation onto glass fiber filters positioned in disposable syringes. The DNA of phages is released by a 50% formamide/4 M sodium perchlorate solution, washed in filter-bound form, eluted with a small volume of low-salt buffer or water, and finally recovered by centrifugation. Comparison of the DNA isolated by this method with that obtained by two conventional procedures reveals both a similar recovery and a similar suitability for restriction enzyme digestion and subcloning.
Jakobi
R
R
Institute of Experimental Pathology, German Cancer Research Center, Heidelberg.
Wiemann
S
S
Pyerin
W
W
D010582:Bacteriophage lambda / Q000032:analysis* / Q000235:genetics; D011232:Chemical Precipitation; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D004926:Escherichia coli / Q000032:analysis; D005374:Filtration / Q000295:instrumentation; D005898:Glass
D004269:DNA, Bacterial; D004279:DNA, Viral