I extracted total RNA from my samples and I ran an agarose gel 2%. You will see two bands on the top of the gel, and one strong band on the bottom, which is my loading dye (bromophenol blue), and something else, probably. I looked at other gels, and people from my lab never saw a band (on the bottom) so strong like that.
Hi Aletheia,
I think the gel looks fine...even in our plant samples we see the lowest band on a regular basis...the quality of RNA also llooks fine...This band of concern basically contains small RNA fragments (also as a consequences of shearing/ degradation) and I feel your small RNA fragments have been efficiently precipitated in this experiemt....as quality looks fine you may go ahead with the exepriment
bye
Thank you so much Ajay, I tought it could be something bad like inhibitors, and I would have to start over.
Althoug late, I like to say that it would be mostly lower MW RNA and mRNA. Usually, the ladderish trail along the lane shows slight shear/degradation. This is emminent in manual isolations. When done with kits, it is not visible much when u follow the exact procedure. But in this case, if used for a RT reaction, it should work good. Also u need to do the DNAase treatment before that.
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