My work is on microRNA on alfalfa. I need to purify and isolate mRNA from Alfalfa extracted RNA using Dynabead purification kit. The protocol given was for 75ug of RNA. I don't need that much amount. I just need 250ng of mRNA in later stages. I am purifying mRNA to use it for 5'RACE to verify if miR408 cleaves on those sites of its targets genes or not?
I tried purifying mRNA using Dynabead purification kit using 5ug of RNA. I wanted to know if we can use nanodrop to check its concentration, yes then should it have same peaks as total RNA or not. I tried nanodrop but there is so peak as we see in total RNA
Please fellow researches, have some comments :).
I would try again but follow the kit guidelines for using 75 ug. Even if you don't need that much, it is likely that the kit works better with the suggested amount. For example, you might need to saturate binding to the beads. If you really want to use less, scale down proportionally.
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