Small RNA-seq
first, polyacrylamide gel electrophoresis (PAGE) purification of the RNA bands corresponding to 1830-nt size fractionation was performed to recover small RNAs; second, ligation of the 5p and 3p adapters to the RNA occurred in two steps, each followed by PAGE purification; third, RT-PCR amplification was performed to generate a cDNA colony template library and then purified by PAGE; and fourth, the template was analyzed using an Agilent 2100 Bioanalyzer (Agilent RNA 6000 Nano Kit) and qPCR was performed to produce a qualified library for sequencing.