We have tried to isolate RNA from tissue sample obtained from a patient with lumbar disc degeneration. We have used a piece of tissue from the surgical sample. We did not get a good quantity of RNA from this. Is there a specific area of disc tissue which can be used for gene expression study? We have used invitrogen RNA isolation kit for this purpose.
I don't have experience with disc tissue, but we have had good results with other difficult, fibrous tissues (cardiac and skeletal muscle) by freezing the tissue on dry ice or in liquid nitrogen then pulverizing it mechanically in a mylar bag prior to RNA extraction. Another method for difficult tissues is to use a bead mill, which we use for hair extractions. Of course, disc tissues have relatively few cells, so you may also want to increase the amount of tissue you extract.
Hello Sibin,
I agree with Richard, i think you need to increase the amount of tissue, but i also recommend you should work very fast to avoid a possibly RNA degradation. Consider use RNAlater or nitrogen liquid for storing samples.
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