Does one need to cleave off the poly A before ligation

Dr. Randal W. Giroux randyg at puccini.crl.umn.edu
Fri Sep 27 09:45:09 EST 1996


We are in the process of making vectors to produce transgenic plants. The
cDNA clones we wish to use have polyA tails on them (as should most
cDNAs). Now if we are going to clone these into a vector between a
promotor/intron complex and a NOS terminator sequence, do we need to
modify the cDNA to remove the polyA from the sequence (ie through PCR). I
have read no ligation methods that suggest this is important but in terms
of expression of the gene product I wonder if there would be significant
problems if the message has two strings of poly A (ie. coding
region-polyA-partial nos term sequence-polyA). I'm stumped!

If any one has experience with this type of modification on expression or
if anyone has some evidence that this is not an important consdieration
please email me directly or reply to this message.

Thanks in advance

Randy



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