synthesis of long cDNA - (Sep/05/2006 )
hi
I am wondering if any of you have a sugestion for a very good Reverse Transcriptase.
I am trying to make a relative long cDNA ~8 kb
also do you know what are the RT conditions for generation of such a long cDNA? do I need to longer the extantion time?
is it possible to do PCR on such a long product (if I'll get one...)?
thanks
keren
if this is for real-time, do you need the entire cDNA length? can you not pick primers that will amplify one end or the other, so if you don't get the whole thing it will be OK? (for example, use oligodT primers and amplify a portion near the polyA tail)
however, you should be able to get long cDNA with the right protocol. I would check Ambion's website
I would also do a google search, something like "long cDNA reverse transcription method" or something similar to pull up protocols...or "synthesis of long cDNA", something like that
however, you should be able to get long cDNA with the right protocol. I would check Ambion's website
I would also do a google search, something like "long cDNA reverse transcription method" or something similar to pull up protocols...or "synthesis of long cDNA", something like that
hi aimikins
I'm trying to prove that there are two ORF on the same mRNA and they are ~5 kb away from each other with no known introns in this long sequence.
I thought to do RT with primer from the downstream ORF and then PCR with primers from both ORF
if you have any more suggestions I'll be happy but I'll try to google it anyway
thanks
keren