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.
More information about the Methods
mailing list