Hello, everyone! I have a big doubt about a recipe for a polyacrylamide gel for RNA... it says "add 5.6 mL of 40% acrylamide (acryl:bis acryl = 19:1) to the gel mix for a 15% polyacrylamide gel". I don't understand what that 40% means if I have the 19:1 relation. Should I put 20g of the 19:1 polyacrylamide mix in 50 ml and then take 5.6 ml of that solution for the gel mix?
I run a 1% agarose gel for my samples of plant RNA in 2 occasions: one with my sample obtained from using a kit (RNAqueous kit from Ambion), and the other one, obtained by using the standard protocol with trizol.
My problem is that I could see both bands from rRNA's subunits in the 1st gel, but only one band in the 2nd one. Don't know what that only band would be for, and why there aren't 2!
I obtain much more yield with the 2nd method, so it's the one I need to develop since I need a pretty big amount of RNA for sequenciation.
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