Your Favorite dsDNA Sequencing Method

fzhedric at hamlet.ucdavis.edu fzhedric at hamlet.ucdavis.edu
Mon Aug 10 19:05:23 EST 1992


Hello,
	I have recently isolated some positive clones from a directionally
constructed cDNA lambda ZAP library.  I am mainly interested in the 5' end of
the molecule as I have already sequenced 80% from the 3' side.  But because
the phagemid is SK+ and ssDNA rescue produces a ssDNA that can only be
sequenced from the 3' end, I am considering dsDNA sequencing.  Because I
have never done dsDNA sequencing before (due to complaints from fellow
graduate students that it produces too many GC compression artifacts),
I am rather ignorant of current state of the art techniques for dsDNA
sequencing.  So... I am wondering if anyone out there would be so kind
as to post their favorite dsDNA sequencing method, including how they prepared
their template and what kind of results they normally get.  I would really
appreciate any advice (including 33-P vs 35-S vs 32-P experience).

	
					Thanks,
					Mike Oda
					fzhedric at hamlet.ucdavis.edu

==============================================================================
          DISCLAIMER NOTICE		 |  Mike Oda
My boss doesn't known what I am doing... |  Biochemistry Grad. Student
And neither do I.			 |  fzhedric at hamlet.ucdavis.edu
==============================================================================



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