Hello RG community,
I want to determine the effect of miRNAs on the 3´UTR of retinoid X receptor alpha mRNA and therefore want to analyze its mRNA level in HepG2 cells. I want to go and check for the mRNA by normal PCR and later Real-Time PCR. However, I am looking for the correct primer pairs but cannot find it. There are 3 transcript variants of RXRA displayed in NCBI. Is it so that I do need to check for those 3 mRNAs (?) and who has analyzed the expression of RXRA in human cells and could provide these primer pairs?
Thanks for your help.
It could be that your miRNAs have differential impact on the three transcript variants. In this case, you would have to quantify the three transcripts separately to determine if any of them is affected. Try the methods section in the following papers for primers. You could blast them against the three transcript variants to determine which they pick up.
https://academic.oup.com/jcem/article/89/11/5851/2844807
Article Time-Resolved Expression Profiling of the Nuclear Receptor S...
you can use one of these primers from primerbank
https://pga.mgh.harvard.edu/cgi-bin/primerbank/new_search2.cgi
Thanks Julie, thanks Abrar,
appreciate your ideas.
I was lucky to find the RXRalpha Fw and Rv primers in several papers with authors performing RT PCR for RXRA quantification.
why you dont design them by your self using Oligo7 primer design tool , if you want I could design them for you ?
Hi Milad,
thanks for your offer. We found RXR fw and rv primer sequences in recently published RT-PCR papers. Primers were ordered and PCR performed. Although the PCR product is faint it worked out quite fine.
All the best
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