Purification by ammonium sulfate precipitation of bacteriophage lambda gt11 DNA for restriction analysis of cloned cDNA inserts.
A rapid and efficient method to purify lambda gt11 DNA is described. This technique involves precipitation of intact bacteriophage particles with ammonium sulfate, followed by phage lysis with sodium dodecyl sulfate, proteinase K, and alkaline treatment. The quality of DNA for subsequent restriction analysis, infectivity, subcloning, and radiolabeling is comparable to that isolated by cesium chloride banding or ion exchange chromatography. The yield of the phage DNA is, however, two to eight times higher than that obtained by other conventional methods of lambda gt11 purification. Furthermore the time required to process the bacteriophage lysate is approximately 2 h and therefore more rapid than other currently used methods.
Ziai
M R
MR
Department of Microbiology and Immunology, New York Medical College, Valhalla 10595.
Giordano
A
A
Armandola
E
E
Ferrone
S
S
D000645:Ammonium Sulfate; D001433:Bacteriolysis; D010582:Bacteriophage lambda / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D004262:DNA Restriction Enzymes / Q000032:analysis; D004251:DNA Transposable Elements; D004274:DNA, Recombinant / Q000032:analysis; D004279:DNA, Viral / Q000302:isolation & purification*; D010957:Plasmids
D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D004279:DNA, Viral; D004247:DNA; D004262:DNA Restriction Enzymes; D000645:Ammonium Sulfate