RNA electrophoresis - (Jul/06/2002 )
RNA was extracted from the brain tissue in order to northern blot, then electrophoresis was explored to exam the integrality of RNA. when the RNA was treated with formamide, formaldahydel and loading dye (xylene cyanole, bromophenol blue, glycerol DEPC water) to degeneration, no bands was showed. When RNA was only treated with loading dye, 3 bands(5s, 18s, 28s) could be observed clearly. There's some problems with formamide, formaldahydel ? How can I solve this problem.
help! helP! sos!sos!
Try using 4.0 ul of 5X MAE, 3.5 ul of (37%) formaldehyde, 10.0 ul of formamide, and 1.0 ul of EtBr (5 mg/ul) per tube (with about 4.5 ul of your RNA sample. Incubate at 55 degrees C for 15 minutes then add 6X loading dye to each sample and load onto your gel.
Are you storing your samples on ice while mixing everything? How much of the Formaldehyde and formamide are you using? Are you running the samples on a FA gel?