Produção Nacional
Rapid silver staining and recovery of PCR products separated on polyacrylamide gels.
1994; National Institutes of Health; Volume: 17; Issue: 5 Linguagem: Inglês
Autores
Sanguinetti Cj, Emmanuel Dias‐Neto, Simpson Aj,
Tópico(s)Molecular Biology Techniques and Applications
ResumoA 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.
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