RNA Extraction Method - (Oct/03/2006 )
i currently use Promega's SV Total RNA extraction kit for extracting pure total RNA to be used in PCR and real-time PCR.... does anyone else know of a cheaper/better alternative?
I extract total RNA using TRIZOL Reagent following the standard protocol (Invitrogen) which leaves you with about 100 microliters of RNA...to further purify it I use RNeasy MinElute CleanUp (Qiagen) which leaves you with about 20 microliters. So using a kit (GeneAmp; Applied Biosystems) to make cDNA, you get 20 microliters cDNA per 1 microliter RNA used (=200microliters cDNA possible).
Hope this helps
I do (almost) the same thing: Trizol isolation followed by clean up with the RNeasy Mini Kit.