Related Experiment Videos
Rapid silver staining and recovery of PCR products separated on polyacrylamide gels
C J Sanguinetti1, E Dias Neto, A J Simpson
1Centro de Pesquisas René Rachou, FIOCRUZ Belo Horizonte, Minas Gerais, Brazil.
Abstract:
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.