I am currently trying to isolate adipocytes from adipose tissue in juvenile chickens and want to use those adipocytes for RNA isolation. I found several articles and protocols online but they are for human or rodents, and did not work well in chickens (RNA concentration from about 10^6 cells about 13 ng/uL but the 260/280 and 260/230 not good). Can anyone give any suggestions or efficient protocols for me to try? Thank you!
Hi Xiao,
I used to isolate RNA from 3T3-L1 adipocytes using the standard protocol of Trizol reagent followed by treatment with DNase. These cells produce high yield of RNA and the purity is high. From my experience with adipocytes, the 260/280 ratio is 1.8 and higher in 95% of the samples. I also isolated RNA from adipose tissue of rat and mice using the same method and got similar results. I suggest Trizol reagent for isolation followed by treatment with DNase. I don't think it will differs in chicken adipocytes because all samples undergo homogenization and lysis in Trizol.
Hope this helps.
Hi Xiao
I would suggest you to use Qiazol from Qiagen which offers High yields of RNA from fatty tissues
please find enclosed a publication from our group in which we used this reagent to isolate RNA from adipose tissue
good luck
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