Maxam-Gilbert Who?!

Eric Chace Olivares olivares at leland.Stanford.EDU
Wed Nov 17 08:34:46 EST 1999


Jens Tornoe (imjt at pop.dtu.dk) wrote:
: How come you do not use Sanger sequencing? It is a lot easier to do, and you
: ought to be able to design a sequencing primer that would give the same
: result as a Maxam-Gilbert reaction.

I had originally planned to use Sanger (of course) but after reading 10
or so footprinting papers, all of which used M-G, I figured that people
use it because it is the simplest way of obtaining a sequence ladder that
will match up exactly with the footprinting lanes (because you can
sequence the same end labeled probe that you use in the footprinting)

I'm having trouble determining the primer design required to generate such
a ladder.  To perform such a strategy it appears to me that I MUST use a
5' labeled probe for the footprinting, and subsequently the sequencing
primer's 5' end must coincide with the labeled position in the probe.
Did this sentence make sense?  If not please correct me because I'd love
to be able to use Sanger to do this....thanks.

-=Eric

-- 
Eric C. Olivares
Department of Genetics
Stanford University School of Medicine
olivares at stanford.edu
http://www.stanford.edu/~olivares/




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