I need to amplify the human UCP3 gene which is about 9 KBs. One option is to use long-range PCR. If anyone has done it, could you please recommend any specific PCR master mix?
I have already used Phusion High-Fidelity PCR Master Mix of Thermo Scientific. In this regard, initially, I got the results but then I got nonspecific bands even after the optimization of the PCR reactions and all the reagents volumes. even sometimes no results. Now, I'm willing to work with the Promega Long-range PCR master mix, but before investing in such an expensive product I want to know about your suggestions.
If someone wants more information about my experiments, you can reply with specific questions.
Thank you in advance.
You may wish to consider TaKaRa PrimeSTAR GXL DNA polymerase. Kindly refer to this publication: https://www.nature.com/articles/srep05737#citeas
which RTs do you use? I would suggest either invitrogen ssII or promega mmlv.
if you get a good/complete cdna, then the PCR should be easy.
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