A procedure of silver staining for nucleic acids in agarose gels without pretreatment or drying steps.
In this paper, we report a fast, simple, and reproducible staining protocol for nucleic acids in agarose gels with a sensitivity in the order of 10 pg/mm2. It took only three steps: fixation, incubation with silver ions, and development of the gels (total time 50 min). The resulting calibration curves (area vs ng of loaded DNA) after a densitometric scanning of agarose gels stained with this procedure were linear up to 50 ng of double-stranded DNA. We found this method suitable for routine laboratory use and especially appropriate for densitometric analysis due to homogeneous background development. Furthermore, it avoids the pretreatment and/or drying steps proposed by other authors for agarose gels.
Prieto
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Sistema de Prácticas Finales, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Hospital Dr. J. M. Cullen, Santa Fe, Argentina.
Leonardelli
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Zalazar
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FE
D010582:Bacteriophage lambda / Q000235:genetics; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D003720:Densitometry / Q000379:methods; D015247:Deoxyribonuclease HindIII / Q000235:genetics / Q000378:metabolism; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D004996:Ethidium / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D009696:Nucleic Acids / Q000737:chemistry*; D012680:Sensitivity and Specificity; D016622:Silver Staining / Q000379:methods*
D005456:Fluorescent Dyes; D009696:Nucleic Acids; D004247:DNA; D015247:Deoxyribonuclease HindIII; D004996:Ethidium