can i see the RNA that isolate - (Oct/05/2007 )

Dear friends:
I isolate RNA of virus from human sera i used trifast reagent
i run RNA on gel with RNA dye i see faint band & in other lane i see nothing
the question is

if the presence of band indicate that i have rna in this sample
& by the same way did the lane that have no band indicate that i have no RNA isolated in this sample

& did anyone know method for Rna isolation from sera

thank u every body & hope 2 listen to your advise

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I always do viral RNA extraction from different sources using trizol (invotrogen) to do RT-PCR. Sometimes I see absolutely nothing in the gel, but still have good amplification bands.

I don’t know about your method, but using trizol RNA has to be precipitated with alcohol and washed, as the pellet is very small and sometimes invisible, I loosed it very often, and I thought I had not RNA, now I use glycoblue and see blue pellet.

Hope this help

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You can use glycogen from any company for coprecipitation with RNA or RNA to see a bigger pellet. The advantage of glycoblue (from ambion) is that the pellet is blue and it’s very easy to see. It doesn’t interfere with RT-PCR or any reaction.

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thx u 4 ur great advise

but can i measure the rna concn sperctrometrically , i dont know 2 do it at all, can i know it stepwise??

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Mmm, I’m not sure. RNA suspension result very little stained of blue.
I never quantify my RNA, I just check it in a gel.
May be adding same amount of glycoblue to the water used to calibrate the spectrophotometer, but I’m not sure.

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[quote name='f.s' date='Oct 8 2007, 01:09 PM' post='113642']
thx u 4 ur great advise

but can i measure the rna concn sperctrometrically , i dont know 2 do it at all, can i know it stepwise??


you can do it spectrophotometrically without any calibration

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