I am trying to isolate RNA from a high RNase, high fat tissue. I am using the guanidine thiocyanate/CsCl cushion method. After the centrifugation through the cushion, I have a large gelatinous pellet. The protocol I have suggests to resuspend the pellet in a buffer containg N-lauroylsarcosine and EDTA. When I do this, the buffer becomes milky, and the milkyness can not be removed by phenol/chloroform extraction. Further, the milkyness precipitates out when I add ethanol/isopropanol. Has anyone experienced this, and if so, how did you overcome the problem? I am not getting any RNA out of the tissue, and I have read of other protocols in which the ppt occurs, however there are no suggestions as to how to prevent it, or any explanation as to what it might be.
RNA Extraction from high lipidtissue
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