Hi Everybody,
Still questions about PCR for circRNA. I do some review for papers and found many of them use MMLV for DNA systhesis. Well in my lab, we usually use Primescript from TAKARA. and I got mutiple bands after PCR. Could this due to difference enzymes?
Is there any different between these 2 enzymes, besides length, time, temp?
Any reply will be help, many thanks
- @Monica- even primescript contains MMLV. Problem may not be due to this enzyme. Try to check the primers specificity or make some changes in PCR setup (may be in annealing temperature, or concentration of MgCl2).
The PrimeScript RT-PCR Kit is a two-step RT-PCR kit featuring PrimeScript RT, an RNase H Minus MMLV reverse transcriptase enzyme that provides robust reverse transcription of virtually any RNA template. But M-MLV RT is a recombinant DNA polymerase that synthesizes a complementary DNA strand from single-stranded RNA, DNA, or an RNA:DNA hybrid. M-MLV RT lacks DNA endonuclease activity and has a lower RNase H activity. M-MLV RT can be used to synthesize first-strand cDNA up to 7 kb.
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