Electroblotting of double-stranded DNA for hybridization experiments: DNA transfer is complete within 10 minutes after pulsed-field gel electrophoresis.
A rapid, simple, and efficient method for DNA blotting is presented. This method is characteristic in that DNA is transferred without denaturation from an electrophoretic gel to a membrane in low-salt concentrations and denatured on the membrane after blotting. More than 89% of double-stranded DNAs ranging in size from 75 to 1.9 million bp can simultaneously be transferred from the gel to a positively charged nylon membrane within 10 min. The present "low-salt electroblotting" method is superior to other blotting methods in that it saves time and labor and its high and even transfer efficiency makes it useful for hybridization analysis.
Ishihara
H
H
Division of Chemical Pharmacology, National Institute of Radiological Science, Chiba, Japan.
Shikita
M
M
D015139:Blotting, Southern / Q000295:instrumentation / Q000379:methods*; D002021:Buffers; D002460:Cell Line; D004247:DNA / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008567:Membranes, Artificial; D008970:Molecular Weight; D009757:Nylons; D013997:Time Factors
D002021:Buffers; D008567:Membranes, Artificial; D009757:Nylons; D004247:DNA