Sequencing from chromosomal DNA

Alexander Kraev kraev at bc.biol.ethz.ch
Tue Jun 25 04:05:21 EST 1996


  Gerd Kleiber <gkleiber at med.de> wrote:

>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<

I think I did not understand something here. Is "linear sequencing"
a non-cycle sequencing (aka T7 polymerase)? IF so, how it can work with 
chromosomal DNA UNLESS it is amplified first? DIRECT sequencing of 
bacterial chromosomal DNA should be possible with cycle sequencing with 
amounts of >1 fmol, but I am not sure if it is possible with fluorescent 
labelling, unless you use some sort of trick to increase sensitivity
about 10-fold. Am I missing something?


Alexander Kraev, PhD                     Internet:    
kraev at bc.biol.ethz.ch
> Lab.of Biochemistry III                            Phone:   0041-1-632-31-47
> Swiss Federal Inst. Of Technology           FAX:      0041-1-632-12-13
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> CH-8092 Zurich                                     /Sasha/kraev.html





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