I have a question.....i have made a agarose gel of bacterial RNA and i have obtained a smear....what's happened? the RNA is 588mer. Moreover, i have done a Real Time PCR of these RNA without amplicon.
Dear Emilia,
If its a smear it means your RNA is not of good quality it seems like its degraded. You should treat all the things like Tips vials everything with DEPC water before isolating RNA DEPC will deactivate RNase so you will get good quality of RNA than you can go for PCR..
To add to Ishan´s submission, its very important to autoclave the tips and collection tubes used in RNA isolation. During the isolation process, it is advisable to change gloves often in order to reduce the chances of contamination of your samples with RNases.
Finally, preparing buffers with autoclaved water or DEPC water also increase the chances of getting quality RNA :
RNA is also sensitive for voltage of electrophoresis. Once I ran it at about 100 volts and I obtained a smear as well.
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