pcr sequencing

Lisa Bibbs at scripps.edu
Mon Feb 27 19:08:10 EST 1995


In article <3ifjfl$ktn at infoserv.rug.ac.be>, dieter.deforce at rug.ac.be wrote:

> Hello,
> 
> I would like to sequence a PCR product. I would use cycle sequencing on
an ABI 
> 373 Perkin Elmer. Is it necessary to clean the first PCR product to continue 
> with the cycle sequencing? Do I first have to remove the primers of the first 
> reaction, the dNTP's? Does anyone have a good protocol?
This depends on whether you are using dye primers or dye terminators.  If
it is primers thank no you don't have to clean it.

Terminators?  Obviously you have to clean it up.  We use an Amicon 100
Micro filter to do it.  Wash it with water.  If the product is clean, you
should get clean data.
Lisa Bibbs



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