High fidelity RT-PCR enzymes - looking for choices... (Feb/23/2005 )
Hi !
I'm looking for high fidelity RT PCR enzymes with proofreading capacity. What is your enzyme of choice?
Thanks in advance
I'm looking for high fidelity RT PCR enzymes with proofreading capacity. What is your enzyme of choice?
Thanks in advance
I am working with Sensiscript RT Kit and ProofStart Polymerase in two-steps RT-PCR and I´m getting good results with them. Both are fron Qiagen. In Qiagen web you can find its characteristics.
I hope it helps.
Superscript III One-Step RT-PCR System with High Fidelity Platinum Taq from Invitrogen rocks.
I had good results with the Titan-One-Tube HiFi RT-PCRT kit from Roche
mike
Hmmm as far as I know there is no such thing as a "proofreading" RNA dependent DNA polymerase. At least that would not be of any advantage to the retro-virus producing it. However some companies may have made blends that will work with proofreading activity?
Anyways - my suggestion is to use any MLV based RNase H negative RT enzyme - e.g. SuperScript from Invitrogen.
If you need to do a downstream PCR for cloning then use any of the High Fidelity blends around. My suggestion is a new Novagen blend called "KOD Polymerase", which is the most amazing blend I have seen around. First of all the fidelity is amazing, second of all the processivity is just out of this world - 10kb can be done with an elongation step of less than 1 min!!! Also this blend contains antibodies to the two enzymes, so the mix will work as a "HotStart".
Good luck!
I think the proofreading refers to the DNA-dependent DNA-polymerase part of the RT-PCR kits, not to the reverse transcriptase.....
mike