Hi,
I am trying to put together a list of the methods used to determine how much mRNA you have after RNA extraction. My plan is to compare total RNA vs. mRNA amounts before and after rRNA degradation, specifically in Bacteria.
The most common methods I have seen include:
1. RT-qPCR with house keeping genes as targets
2. Bioanalzyer
3. nanodrop/qubit
What is your preferred method for the most accurate quantification of mRNA concentration? Also, if you use RT-qPCR, what gene do you target to get an overall estimate of all mRNA?
Thanks!
Morgan
I didn't understand your question. Frankly speaking, I have problem with your question.
How do you want to separate mRNA and total RNA in prokaryotic cells?
In best of my knowledge eukaryotic cells has poly A on their mRNA so you can separate them from total RNA but in pro, I don't know.
Let me know your idea
Regards
Alireza
Would you say ARN total? If It is so that is necesary do to quantification to know how much product cDNA get in RT-PCR
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