i have isolated RNA from phloem sap of plant. the nano drop reading is good but when i run that RNA into formaldehyde gel or even Agarose gel it was seems like run towards -ve electrode. but marker loaded in gel is running in right direction.
There is no way I can imagine that RNA would be positively charged. I would be sure that you have swapped the positions of the electrodes (that happened to me a couple of times) but then, the marker would be running in that direction too.
Really weird situation. Don't know what to say. I don't believe that the isolates are bad if nanodrop results are ok. Try to repeat just the electrophoresis.
RNA is always negatively charged so no way it can runs across -ve electrodes direction. you might have interchanged the plugs of the electrodes or may be you added something in the solution else it will move towards +ve electrodes. so pls recheck!
Are you sure you isolated RNA? It sounds a little like genomic DNA which might stuck in the wells due to its size and looks like reversed electrophoresis
ya i analysed the situation lots of protein contamination is there because i directly isolate it from phloem sap, i gave DNAse treatment to them but still protein is there.
now i purified it with coloum n made cDNA from it its working fine.
I have the same situation going on these days. We are working on single stranded 'C' rich DNAs. In the agarose gel, the DNAs are moving toward the cathode, while dye, and polythymine ss dna are moving in the right anode direction. I can't think of any reason. We repeated 3 days. Same result.
Any clue what is going on?
Similar situation here...prepared RNA probes, purified via columns, ran on denaturing gel, actually seeing two bands, darker bank running in opposite direction while faint band in correct direction. 260/230 ratio were less than 1. Can only imagine protein contamination in bound form with RNA is dragging majority of RNA in the opposite direction. Any comments? thanks
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