Dear Sir/ Madam,
I am designing a primer for microRNA reverse transcription. But some microRNAs like hsp-mir-21-5p has a strong secondary structure. It forms a stable hairpin.
Can anyone please tell me whether the secondary structure of the microRNA(especially stable hairpin structure) will influence the reverse transcription?
If possible, please recommend some good RTase applicable in reverse transcription with severe secondary structure!
Thanks!
Binbin
Hey,
AMV reverse transcriptase is actually reducing the hairpin structures.
Article [33] First-strand cDNA synthesis primed with oligo(dT)
Even if secondary structure of some miRNAs have low MFEI or high stability, but a mature miRNA to get formed i think precursors will surely be processed. You need not worry about stability of miRNA precursors and should go ahead with stem loop RT primer designing.
- Badges
- Science method