Direct sequencing of lambda phage dna

Guilherme Co. Oliveira guilher at tam2000.tamu.edu
Mon Nov 7 14:14:22 EST 1994


	I have done this by extracting DNA according to the protocol in
Maniatis (Qiagen preps also work well), and using the Sequitherm (Epicenter
Technologies) sequencing kit.  This is a PCR sequencing procedure, and the
modifications I use is that in the first cycle I denature the virus particle
and the DNA at 95C for 5 min.  I use an annealing step for 30s and 1.5min. of
elongation.  Also I add 1ug of DNA and 3pmols of primer.  Doesn't work as well
as double stranded plasmid, but you get results.  Good luck.



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