Cheap and best method for full length m-RNA purification - (May/26/2008 )
I am running some invitro transcription experiments.
Although most of my RNA is going to be mRNA, I am interested in only analyzing full length mRNA later.
Can anyone suggest a reliable and cheap method to isolate only full length mRNA?
There is a kit from Epicentre called "mRNA Only" which selectively digests rRNAs in you total RNA sample. I am currently in the phase of testing it and it seems quite promising, at least you don't observe any rRNA after the digestion and I can catch specific transcripts afterwards from the mRNA prep. But at the moment I don't know if all transcripts are full length or not, i think there is no way of knowing it...
One thing you can for sure try: work RNAase free and minimize the risk of partially degraded mRNA:) You might also want to make sure that your preps are free of metal ions and stuff that may fragment mRNA.