title: N2 ints-11 RNAi long scRNAs
design: 2ug total RNA was 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.