I am having issues extracting RNA from adipose tissue. Currently, I am using the miRNeasy Mini Kit from Qiagen following their protocol. The result is a low concentration and an A260/A280 of ~1.8-1.9. I then run the sample on a 1% agarose /1X TAE/1% Bleach RNA Gel. The bands are always smeared. I treat everything with DPEC treated water, change my gloves regularly, and use RNase Zap liberally. I’ll include an image of one of my gels later today for those that are interested.
I usually do my RNA extractions with Trizol, however, some common problems with RNA extraction from adipose tissue are poor homogenization and a high content of fat.
I would make sure that the tissue is correctly homogenized before proceeding with the next step. Also, It might be wise to add an extra step after homogenization/before chloroform. I centrifuge at 12,000g 10 min at 4ºC to obtain two phases, the first one with the fat and the second one with the lysis buffer. I transfer the liquid phase into a clean tube and continue the extraction as usual.
Hope it helps!
the main point for RNA extraction is homogenization. for this goal, you could try a different method like Usain homogenizer or etc. I think by keeping this in your mind you get better results
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