I have used Trizol based liquid nitrogen method but not getting a good yield. I need it for cDNA preparation and qRT-PCR experiments.
Any suggestions are welcome. If possible share the protocols.
Thanks in advance.
You need to physically break the cells with bead beating using tiny glass beads. We use a "Fastprep" instrument from MP Bio and the corresponding blue kit/matrix (http://www.mpbio.com/product.php?pid=116004500&country=222); works very well for all mycobacteria.
In my opinion, a good kit of total RNA isolation is "RNeasy Mini Kit" from Qiagen. I often use it for RNA isolation from lymphocytes and epithelial cells.
https://www.qiagen.com/us/shop/sample-technologies/rna/total-rna/rneasy-mini-kit/#orderinginformation
By the way, I've find on Qiagen website a specific supplementary protocol for bacteria. So, I'm sending it to you and I hope it may be usuful for your problem.
Regards
In our lab, we regularly use for isolation of RNA from M.smegmatis combination of sonication + phenol/chloroform extraction.
The RNA is after that in good quality, we can also make NGS with it.
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