HELP!! Artificial poly(A)!!

JongSub Lee jongslee at cd4680.snu.ac.kr
Mon Nov 28 06:56:54 EST 1994


Hi everyone..

I stuck in a great problem. Please help me.
I got a cDNA clone having 9 poly(A) residues on 3' end. Its size is
7.4kb. It was cloned into pUC18/SmaI. 
But there are few poly(A) residues on 3' end, I have to add poly(A)
on its 3'end. Unfortunately, Its size is not small (7.4kb), there are
many restriction sites in it. All the restriction sites in the pUC18 
multiple cloning site are located in 7.4kb cDNA insert.
So, if I cut a site and gonna use TdT, the cDNA insert will be break
into some pieces.

Oh.. God...
What should I do ?

If you have brilliant idea and reliable protocol on this,
please let me know.

Francisce Xavier





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