The "Race for 5'ends

Matthias Zeiner mzei at sun0.urz.uni-heidelberg.de
Fri Mar 18 10:58:27 EST 1994


----- Forwarded message begins here -----
From: kumar  <KUMAR at JII.AFRC.AC.UK>
Fri, 18 Mar 94 13:41 GMT
To: MZEI <MZEI at sun0.urz.uni-heidelberg.de>
Subject: Re: The "RACE" for 5'ends

Hi,
I would like to know whether your 5' truncated cDNA was isolated from a cDNA
library? If that is the case, just forget 5'RACE but try to amplify 5' end by
using primers based on vector and 5'end sequence with total DNA from the 
library as template.
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Thanks for your suggestion but I exactly tried this without success. I used 
lambda gt11 universal primers and two nested gene-specific primers 
located near the known 5'end. All I got was a smear of products but no 
additional 5'information, probably because the cDNA library is full of 
truncated clones. 
Maybe it could be worth-while to try again using a third nested primer even 
closer to the known 5'end to enhance specificity and to exclude shorter 
cDNA's. 

Matthias



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