Sequencing from chromosomal DNA

Gerd Kleiber gkleiber at med.de
Mon Jun 24 16:37:53 EST 1996


Rafael Maldonado wrote:
> 
> Hi netters,
> 
> Does anybody know if it is possible sequencing directly from bacterial
> choromosome preps? Do those cycle sequencing systems work in this case?
> 
> We have got the sequence of most of our gene, and we would like to
> sequence the rest without cloning it. There are only 160 bp missing!!
> 
> Any comments appreciated.
> 
> Rafa

Hi Rafa,
Of course. Just needs DNA which is enough clear... usea commercial kit, 
I´d recommend Quiagen or someting similar. Cycle sequencing and linear 
sequencing both work. We do routinely Minipreps ans mentioned above, then 
sequence them with the new ABI cycle seq. kit in a 
one-cup-one-PCR-reaction. Works up to 800 bases.
Ask for more if you want.
Sandra




> 
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