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RNA Extraction from high lipidtissue


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#1 anonymous

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Posted 08 February 2001 - 10:00 PM

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.




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