Hello everyone,
I want to measure the expression change of some mRNA, miRNA, and LncRNA at once under a specific condition.
Since I am going to do relative quantification, is there any RNA extraction reagent that can extract all RNA molecules simultaneously?
I am willing to hear if you have other solutions rather than a reagent for all molecules.
Thanks for your help.
Hello,
It depends on your lab.
If you have money I would definetaly suggest you to use mirVana total RNA isolation kit.
If you dont have that much money, you can use Trizol Reagent.
For miRNA extraction, either isolate miRNA using isolation kit and then cDNA synthesis and the using miRNA specific primers go for PCR.
Or you can isolate total RNA using Trizol and then using miRCURRY RT Kit II, convert all miRNA present in total RNA into miRNA-specific cDNA. That cDNA can be directly for SYBr based qRT-PCR with miRNA primers.
I am not well versed with other LnRNA isolation.
Thank you dear Enes Yağız Akdaş ,
The mirVana is a little costly. Does trizol give at least 30 ng of miRNA?
Thank you for your answer dear Saurabh Mandal,
However, I need miRNA, LncRNA, and mRNA together. I wonder if I can extract them separately and then compare their expressions.
Then, I think you need to buy isolation kit which may come with different pore size filters to get all three together in one go.
There are also Qiagen kits for DNA/RNA isolation from the same sample. I have previous experience with the AllPrep DNA/RNA/miRNA Universal Kit, which worked well for DNA, long RNAs (lncRNA, mRNAs…) and miRNAs simultaneous extraction. Alternatively, you could use a total RNA extraction kit and then use separate RT-qPCR kits and primers for miRNAs and mRNAs/lncRNAs.
Saurabh Mandal and Perla Pucci many thanks for your responses.
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