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mhughes at mbcf.stjude.org mhughes at mbcf.stjude.org
Wed Aug 30 16:57:35 EST 1995


In article <DE27w7.6JK at news.nsw.CSIRO.AU>, D.Adelson at prospect.anprod.csiro.au (David Adelson) writes:
> Greetings Netters,
> 
> We are about to start looking for some sheep tissue 
> specific ESTs, using a lambda ZAP cDNA library.
> 
> Is it possible/better to use PCR to amplify the insert
> regions of individual clones and then cycle sequence
> the PCR product directly?  We are looking to avoid having
> to do the in vivo excision, plating and miniprepping to
> get template for the sequencer.
> 


Dave,
	We do RT-PCR of flu virus RNA all the time and get plenty of product
for cycle sequncing.  The only problem is you can't get very good sequence
within 30 or so bases of the termini.  If you need that data however, its easy
enough to ligate any PCR products that look "interesting" into a T-vector and
complete the sequence from there.
	Hope this helps.
Mark

Mark Hughes 
St. Jude Childeren's Research Hospital
Dept. Virology
Memphis,TN
 



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