2 replies to this topic

[MichaelM]

Hi everybody,
I just wanted to know what's your protocol for extracting this kind of tissue? I use Trizol with an extra chloroform extraction. I haven't gotten good results from this and have never gotten an RNA pellet. Though I don't get an RNA pellet I still continue until the end and most of the time get a 260:280 ratio above 2.1 (usually around 4-6). I tested for contamination, but I always get good results for other tissue (i.e. liver). I'm just having a hard time on this one. If you can help me thanks!

[gfischer]

In adipose tissue, you may have some contaminating fat in your sample.  According to Invitrogen's protocol after homogenization, you can remove insoluble material by centrifuging at 12,000g for 10 minutes at 2-8C.  In adipose tissue, there should be a layer of excess fat on top, then the supernatant with RNA, then a pellet containing other insoluble materials, like cell membranes and high molecular weight DNA.  remove the fat layer, then use the supernatant for the extraction.

[MichaelM]

i forgot to mention that I do remove the fat layer. Still no pellet. I also use about .05-1 g of tissue. Does the tissue size for fat matter?