title: TFIIS EV germnuclei nucleoplasm rep 1 scRNAs
design: 500ng total RNA were treated with 2l Quick CIP (NEB) in a total volume  of 20l for 90 min at 37C. RNA was phenol-chloroform extracted, and  precipitated at -20C overnight with 1/10 vol 3M AcNa, and 3 vol 100% EtOH  and 1 l glycogen. The next day, RNA was resuspended and treated with 7.5  units (1.5 l) RNA 5 pyrophosphohydrolase (NEB) for 1 h at 37C in a total  volume of 20l. RNA was extracted and precipitated overnight as described for  CIP treatment. The resulting RNA was used as input for small RNA library  preparation using the TruSeq small RNA kit (Illumina) according to the  manufacturers instructions, except for an increase in the number of PCR  cycles from 11 to 15. An insert size range of 20-70nt was gel purified.