Rapid silver staining and recovery of PCR products separated on polyacrylamide gels.
A rapid silver-staining procedure for DNA fragments in polyacrylamide gels is described. The time required for band detection is 15 min and the limit of sensitivity 3 pg/mm2. PCR products subjected to this rapid staining protocol are readily recovered from the gel by excision and elution by incubation at 95 degrees C for 20 min. Bands of up to 3 kb have been recovered and reamplified from either recently prepared or dried gels. The rapid staining protocol significantly decreases the processing time required for silver-stained polyacrylamide gels, which is of particular importance in diagnostic situations. The recovery protocol allows individual bands from complex mixtures to be easily recovered for sequencing or probe preparation.
Centro de Pesquisas René Rachou, FIOCRUZ Belo Horizonte, Minas Gerais, Brazil.
C J
CJ
Sanguinetti
E
E
Dias Neto
A J
AJ
Simpson
D000818:Animals; D001483:Base Sequence; D018509:DNA, Helminth / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012550:Schistosoma mansoni / Q000235:genetics; D016622:Silver Staining
D018509:DNA, Helminth