parn-1 small RNA CIP-Rpph
~5 ug of total RNA were pretreated with 4ul Quick CIP (NEB) for 90 min at 37C in a total volume of 40ul, then incubated for 2 min at 80C. The RNA was phenol-chloform extracted and ethanol/AcNA precipitated, resuspended in water and treated with 1.5 ul RNA-pyrophosphohydrolase (Rpph, NEB) in a total volume of 20 ul for 1h at 37C.  The RNA was phenol-chloroform extracted, ethanol/AcNA precipitated overnight, resuspended in water and used as an input for TruSeq small RNA library preparation kit from Illumina according to manufacturer's instructions.