Does anybody know which is the best method to obtain a small RNA-enriched RNA preparation from plants? TRIzol, Quiagen, some modification of them, etc.
Hi Pedro
There are several kits available to enrich the small RNA fraction...from SIGMA, Roche, Qiagen etc......However you can also use some established methods in literature that doesnot depend upon the kits.....Such methods use TRIZOL for total RNA isolation, no 70% ethanol washing step...and finally selective precipotation of large RNAs from the total RNA...and what left is small RNA fraction.......
For isolating 20-24 nt long RNAs...the best way is to run a 15% denaturing gel and isolate the fraction
bye
Peter, what Ajay said is true. You can just Trizol your samples. Howeverr, if you can get your hands onto an Ambion kit (I think now from Invitrogen) it may make your life easier. Good to hear from you !!!!
Beware! most kits for purifying total RNA lose the small-RNA fraction!!! You must buy specific kits that retain these RNAs, like MirVana from Ambion, and other brands. These kits are designed to purify microRNAs but also retain all sorts of small RNAs
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