I want to study few long non coding RNA and need to know its localization through qRT-PCR. Can anyone suggest me any method to fractionize RNA? After putting up qRT-PCR, what controls should I use inorder to normalize it?
Francisco J. Enguita
Hi Prit
To fractionate lncRNAs from nuclei you can use whatever protocol for the isolation of nuclei (there are a few around for doing Chip, for instance), and then extract the RNA with your favorite kit or home-made protocol in the nuclei and cytoplasmic fractions. If you want to use a normalizer for qPCR experiments I would suggest you to use any snoRNA for the nuclear lncRNAs and a protein coding transcript for the cytoplasmic ones (GAPDH or others).
Best and good luck
Paco
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