Rapid silver staining and recovery of PCR products separated on polyacrylamide gels.
Rapid silver staining and recovery of PCR products separated on polyacrylamide gels.
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发表时间：
1994-11
期刊：
影响因子：
2.7
通讯作者：
CJ Sanguinetti;E. N. Dias;AJ Simpson
中科院分区：
文献类型：
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作者：
CJ Sanguinetti;E. N. Dias;AJ Simpson
本文介绍了一种在聚丙烯酰胺凝胶中快速银染DNA片段的方法。条带检测所需时间为15 min，灵敏度极限为3 pg/mm 2。PCR产物进行这种快速染色协议是很容易从凝胶回收切除和洗脱在95 ℃下孵育20分钟。带高达3 kb已被回收，并从最近制备或干燥的凝胶再扩增。快速染色方案显著减少了银染聚丙烯酰胺凝胶所需的处理时间，这在诊断情况下特别重要。回收方案允许从复杂混合物中容易地回收单个条带用于测序或探针制备。
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.