sequencing PCR products

Karl Fischer tyr-2 at bones.biochem.ualberta.ca
Tue Mar 10 11:31:59 EST 1998


In article <Pine.GSO.3.94.980310095334.12303A-100000 at tin>, "Miss. V.M.
Hodges" <vhodges at hgmp.mrc.ac.uk> wrote:

> has anyone got a good method for manual sequencing of PCR products
> (~500bp). 

Posted and mailed.

Vivien,

We have been doing sequencing of PCR products for a number of years using
the dsDNA cycle sequencing kit from Gibco LifeSciences (no affil). Have
gotten good sequence from a 293 bp PCR fragment without the need to
pre-purify the PCR-generated DNA. Additional benefit is you only need 30
ng/kb of DNA in order to sequence (in your case, 15-20 ng of your 500bp
product should work fine).  

Sorry the suggestion had to be another kit.

Cheers

Karl the hepB guy



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