Ask for help about the RNA purification protocol by phenol/chlorform methods - Ask for help about the RNA purification protocol by phenol/chlorform m (May/11/2009 )

Ask for help about the RNA purification protocol by phenol/chlorform methods

I just start to work on the miRNA qRT-PCR fields and try to establish some assay, the method is firstly to add poly (A) at the 3-end of miRNA, then synthesis the first cDNA strand by reverse transcription

After the isolation of total RNA (Trizol methods from invitrogen), add poly (A) at the 3-end of miRNA, the phenol/chloroform procedure is needed. What is the detailed protocol of phenol/chloroform purification? and after purification, the small RNA should still be included in the total RNA.

Thanks a lot in advance

Veer2

Usually it's just the same like a DNA phenol-chloroform extraction, except you use acid phenol instead of alkaline (we use phenol from QBiogene).
I use make two phenol-steps followed by one chloroform-step.
The precipitation is done with ethanol and Na-acetate. You can also add glycogen which should increase precipitation.

Thanks a lot