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Rapid silver staining and recovery of PCR products separated on polyacrylamide gels.
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1994
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EngineeringComplex MixturesDna AnalysisMolecular BiologyNucleic Acid Amplification TestPcr ProductsPolyacrylamide GelsBioanalysisAnalytical ChemistryMolecular DiagnosticsPolymer ChemistryChromatographyDna SequencingDna ReplicationPolymer ScienceNucleic Acid AmplificationPolymer CharacterizationRapid Silver StainingMicrobiologyRapid Silver-staining ProcedureMedicine
The study describes a rapid silver‑staining procedure for DNA fragments in polyacrylamide gels. The protocol uses rapid silver staining followed by excision and 95 °C incubation for 20 min to recover bands, enabling easy retrieval of individual fragments from complex mixtures for sequencing or probe preparation. The method detects bands in 15 min with a sensitivity of 3 pg/mm², allows recovery and reamplification of fragments up to 3 kb, and markedly reduces processing time, benefiting diagnostic workflows.
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.