I'm working on primary adipocytes with a very high lipid content - to harvest the cells for RNA extraction I have used RLY lysis buffer from the Bioline RNA mini kit. However after extracting RNA from a few of my samples I've gotten a very low RNA yield, which is due to lipids getting stuck on the membrane.
I was wondering if anyone has tried adding Qiazol or trizol onto already lysed cells, then adding chloroform and removing the liquid phase... then continuing with RNA extracting as usual.
I also faced this kind of difficulty in my case, as I dealt with human adipose tissue samples, so what I did just used normally recommended Trizol method with twice the chloroform step, which reduce the total lipid content.
As above. Also, use more TRIzol: double or triple that recommended (so 2 or 3ml per 100mg tissue, for instance).
We work with brain tissues and use QIAGEN RNeasy Lipid Tissue Kit (catalog #74804 for mini kit). Works well and includes QIAzol (I guess it is similar to TRIzol).
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