Hi guys,
I am wondering how to know if miRNA is regulated at transcription level? Should i measure pri-miRNA level instead of mature miRNA level?
Hi Xingchen
Of course !.. You should check for pri-miRNA levels if you want to look at transcriptional regulation. Be careful with the design of qPCR primers to cover the right region of the transcript.
Best
Paco
Thank you sir!
My miRNAs are clustered and located in the intronic region of a host gene. When this intron is cut out from host gene mRNA, is there a poly A tail at 3' end of the intron that we can use oligodT as a primer to do the reverse transcription? I looked at some papers listing the real-time PCR primers for pri-miRNAs and the 3' primer is not universal primers like what we use for mature miRNAs. So i am wondering should i design these pri-miRNA primers as a regular gene? Or still i need to add poly A tail and use a specific adaptor to do RT and then a universal primer to do the real-time PCR.
Thank you!
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