dsDNA sequencing protocol

[Andreas Buhr]

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