PCR product sequencing

nsaunders at molbiol.ox.ac.uk nsaunders at molbiol.ox.ac.uk
Wed Sep 18 16:48:52 EST 1996


I've used the Sequenase kit and manual sequencing to directly sequence PCR 
products of 400bp or so quite successfully. For a really clean PCR product, 
it's possible to use the PCR mix directly as template, though there is some 
background, so you might benefit from a bit of cleaning up. The best results I 
ever had were using a protocol for sequencing PCR products from low melting 
point agarose; this was very clean. The reference is:

Kretz and O'Brien (1993) Methods in Enzymology 218: 72-79

Neil Saunders
Dept of Biochemistry
Oxford University
 



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