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快速银染及聚丙烯酰胺凝胶上分离的PCR产物回收

Rapid silver staining and recovery of PCR products separated on polyacrylamide gels.

作者信息

Sanguinetti C J, Dias Neto E, Simpson A J

机构信息

Centro de Pesquisas René Rachou, FIOCRUZ Belo Horizonte, Minas Gerais, Brazil.

出版信息

Biotechniques. 1994 Nov;17(5):914-21.

PMID:7840973
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.

摘要

本文描述了一种用于聚丙烯酰胺凝胶中DNA片段的快速银染方法。检测条带所需时间为15分钟,灵敏度极限为3 pg/mm²。经过这种快速染色方案处理的PCR产物可通过在95℃温育20分钟后从凝胶中切下并洗脱的方式轻松回收。长达3 kb的条带已从新制备的或干燥的凝胶中回收并重新扩增。该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,这在诊断情况下尤为重要。回收方案允许从复杂混合物中轻松回收单个条带用于测序或探针制备。

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