basic PCR question

[Dr.BoPoWoFo]

Dear Sarah,

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,

Wo

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.
>
>Thanks!
>Sarah

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 -
Germany

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