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 Location: Home > LitDocs > Application Note > Application Note: Recovery and PCR amplification of DNA fragments separated on PhastGel® (for research use only)
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Application Note: Recovery and PCR amplification of DNA fragments separated on PhastGel® (for research use only)

 (18110843.pdf - 38 Kb)


The recovery and PCR amplification of DNA fragments separated on PhastGel ® is described. The method includes a rapid silver staining procedure for DNA fragments in polyacrylamide gels that complements the speed advantages of PhastSystem ™ . The method is significantly faster than previously published methods on which it is based. The time required for band detection is reduced to 15 minutes and the limit of sensitivity is approximately 10 ng/mm 2 . PCR products subjected to this rapid staining protocol are readily recovered from the gel by excision and elution at 95°C for 20 minutes. The recovered PCR products are reamplified by the PCR process. The protocol allows the use of PhastSystem for the routine analysis of PCR products and diagnosis based on PCR amplification. The possibility of recovering silver stained bands may be particularly advantageous in conjunction with SSCP analysis since bands exhibiting altered mobility can be recovered for sequencing.





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