5'RACE of RNA Pol3 transcripts?5RACE Pol2 Pol3
Posted 13 September 2012 - 02:44 AM
I found an interesting Antisense transcript running opposite my gene of interest. So i performed 5'RACE to attempt to map the TSS of this transcript, but after two months, and a variety of different primers i am unable to get a band that corresponds to my transcript. i am getting bands, but upon sequencing, they are not the sequence i was looking for.
There is a U6 snRNA just upstream of this transcript, which is transcribed by RNA polymerase3, so maybe my transcript is also a Pol3 transcript.
So i was wondering is 5'RACE possible with RNA pol3 transcripts? and if not is this the reason that my 5'RACE has failed?
Thanks for your insight.
Posted 13 September 2012 - 10:38 PM
Posted 14 September 2012 - 01:24 AM
I think that this antisense transcript may be endogenously transcribed by RNA pol3, and my question is,
would this 5'RACE protocol work on a RNA pol3 transcript, when it was original designed to amplify the 5' end of a RNA pol2 transcribed transcript.
thanks so much for your help
Edited by jaywilla, 14 September 2012 - 01:24 AM.
Posted 16 September 2012 - 07:44 PM