basic PCR question
novalidaddress at nurfuerspam.de
Tue Jul 12 17:29:16 EST 2005
basically, there is nothing special except that you should choose a "good"
RT enzyme which has enough performance for such long fragments (MMLV is a
good choice if i remember correctly - you also might check AMV) and then
for the PCR make sure you have enough extension time for the fragment. For
Taq, 1 min should be enough but for proofreading Polymerases, you will need
approx 5 mins.
Best of results,
At 10:30 12.07.2005, Sarah LaMere wrote:
>I'm wanting to amplify a 4 kb fragment using rtPCR, and I'm just wondering
>what are the best PCR conditions and/or reagents people have found to
>use? I know it's a very basic question, but I've never attempted to
>amplify a fragment that size with rtPCR, so I just want to get a decent
>idea of the best way to go about it.
Wolfgang Schechinger, Dr. rer. nat.
Med Clinic I, Endocrine Research Lab
Bochum University Hospital "Bergmannsheil"
- the world's first casualty hospital for miners and steel workes -
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