blocking a dna adapter / linker against 5->3 fill in ?
Ralf Sigmund
ralf.sigmund at mpihan.mpg.de
Wed Sep 1 10:57:49 EST 1999
Hi,
I would like to digest cDNA and afterwards ligate it to two different
adapters.
one should look like this:
5'-GNNNNNNNNNNNNNNNNNNACGATGAGTCCTGAG-3'; top linker
3'-TACTCAGGACTCGC-5' bottom linker
!
Now I would like to block the 3'-terminal base of the bottom linker, so that
Taq-Polymerase cannot fill in the 5'-3' direction.
I thought about ordering a oligonucleotide with a 2',3'-didesoxy group in
this position. However this modifaction appears to be very expensive.
Is there a way to block an normal oligo enzymatically ?
TIA
Ralf
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