Long range PCR - What enzymes do you use? - (Aug/21/2006 )
I am trying to decide which long range PCR kit to use. There are so many out there and this will be my first time trying long range PCR. I'll be amplifying anywhere from 10 to 20kb. Does anyone have a favorite that works more often then not?
takara LA works well for me
haven't had much success with Expand (Roche) though
Has anyone used Invitrogen's elongase or BD adventage genomic polymerase? I heard good things about LA takara as well.
i've done few successful exp with Expand 20 kbPLUS PCR System from roche for 15kb.
it was 4years ago... so maybe there are better kits now
Dynazyme EXT from Finnzyme
In our lab, we use Expand long and Expand 20kb plus. i can amplify a PCR product up to 16 kb by using expand long.
i swithced for Eppendorf Triple master recently, not tested up than 8kpb, but at this size, efficiency is very good (better than long range one).
So you may consider it too as the manual says it's ok for episomal up to 50kpb...
so ask for an assay vial.