Small RNA extraction - iRNA (Apr/03/2006 )
Hi, I do the same kind of exp. in my lab; extracting small RNA from silenced Nicotiana benthamiana plants (transgene is silenced). I use Tri Reagent (Molcular Reseach Centre, MRC) to extract the total RNA. To isolate the low molecular weight RNA, I precipitate the total RNA with NaCl and PEG (at final conc. of 0.5 M and 5% respectively). I work nicely in my hand and other labmates who are following my protocol.
according to the page you gave (http://plantpath.unl.edu/llane/text/dsRNAextraction.html) that extraction seems pretty awful so honnestly i won't trust this protocol. I've noticed personnally, that the recommended volume of trizol per cell is not the better. I tend to use more volume than the one recommended n get clean RNA with good quality.
You brought to me an other reason why it was better : i think it's because the pH was to up when using less volume of trizol
Hi everyone, thanks for the answers.
This time i am trying to extract the small RNA following all the advices. I let you know how it goes. thank you very much.