Recently, I have been trying to quantify the expression of miRNA. I have extracted the miRNA. However, I am not clear about using the cDNA synthesis kit usually there are two kits providing by companies the Poly A kit and a stem-loop PCR kit. Please share your knowledge about the miRNA expression analysis, thanks.
Both methods can work, the stem-loop ones can offer a bit more specificity but you will be locked into that chemistry and the kind of probes and primers that come/possible with that kit/method.
I previously used the polyA tailing kit, it offers more flexibility, economic but you are responsible for the forward primer design. Both are possible and good options, I would argue stem loop might be a good start for a starter who doesn't intend to analyze many genes especially if your vendor gives you validated primer probe sets. MicroRNA primer design is a little bit more difficult than coding gene primers that are much longer than your microRNA transcripts. If you are comfortable with taking risks, then polyA can work for you too. Sequence similarity is an issue though.
Can Justin Kiessling Thank you so much for your explanation. I appreciate it, and I've grasped the concept clearly now.
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