dsDNA sequencing protocol

Andreas Buhr Buhr at pki.unibe.ch
Fri Dec 2 06:51:43 EST 1994


Hi All,

try Rob Schuurman and Wilco Keulen's "Modified Protocol for DNA Sequence
Analysis Using Sequenase 2.0" (BioTechniques 1991 Vol.10 No.2 p185) 


There are three changes to the original Sequenase procedure:
- presence of 10% DMSO
- elevated reaction temperature (48 oC)
- chase with Klenow

the result is really FANTASTIC!!! (try it)

For a stronger signal I prefer to use about 0.8 pmol alkai denatured DNA in
a half reaction (single loading).

Andreas

buhr at pki.unibe.ch 



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