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Checking RNA integrity by Denaturing Gel - (Feb/26/2014 )

Dear All, 


I wanted to check the integrity of my RNA sample using agarode gel electrophoresis


I add RNA sample(200 ng) to RNAse free water to form 7 µl + 7 µl loading buffer . then I took this 14 µl and dispense them in each well. To the agarose gel, I used 1.2 gm + 100 ml of 1 X TBE buffer and add 5 µl Roti Saf dye. I adjusted the run to be 1,30 hours at 100 Voltage. 


My results was like in the figure. 


Do you see that this picture indicates a nice working system ? If any one see comments on the protocol, let me know to optimize 

Attached Image

-Mohamed 1984-

The gel looks fine, but the samples don't. These look like genomic DNA samples, with possibly highly degraded RNA present. How were they prepared? You should have two sharp bands, corresponding to the 23S and 16S rRNA (unless you are isolating mRNA, in which case, these will be absent).


You should take some RNA you know it's fine to test "a working system".


You also didn't specify what is your "loading buffer" (which should contain denaturing agent obviously).


Otherwise I agree with phage.