Sequencing G-C rich DNA

Patrick F.H. Lai pfhlai at LOOKSMART.COM
Tue Jul 14 20:24:49 EST 1998


Hayley - The Comet wrote:

> Hi
> Does anyone have a method for sequencing G-C rich cloned DNA?
> The sequencing PCR conditions we are currently using for all > sequen=
cing reactions regardless of nucleotide sequence is :
=0996'C=0910 sec
=0950'C=095 secs
=0960'C=094 mins
> for 25 cycles.
> Please let me know....
..................................................................

Dear Hayley,

I have never done sequencing PCR, but I wonder if 96'C for only 10 sec
would be sufficient for G-C rich sequences to dissociate.
Would a high GC-content dictate a longer time at 96'C as in a regular
PCR ?

Just curious.


Patrick F.H. Lai  < PFHLai at looksmart.com >
Grad Student, U of Toronto

LookSmart =85 or keep looking.
http://www.looksmart.com



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