kim@m44.unm.edu (04/25/91)
I like to run RNA on Glyoxal-DMSO gels. My understanding is that the Glyoxal treatment adds covalent adducts to bases on the RNA (or DNA), preventing annealing, and these adducts are removed by incubation under alkaline conditions (pH > 8.0) before Northern blotting, to allow hybridization. The reaction mixture of Glyoxal, DMSO, etc adds up in volume, however, making it necessary to run a gel of a certain well volume to hold the entire sample. I was wondering if the glyoxalated RNA, since it is covalently modified, can be re-precipitated and dissolved in a small volume of buffer for loading onto a smaller size gel. As long as the pH is neutral or acidic during the precipitation, I would think the glyoxylation should be preserved. Does anyone have any opinions/experience with this kind of treatment? Thanks: Daniel Kim