Small RNA sequencing study of C elegans piRNA biogenesis factors

Identification of defects in piRNAs and piRNA precursors Sequencing of small RNAs (18-30nt). Sample treatments: Secondary siRNAs in C. elegans carry a 5' triphosphate and this has to be removed prior to cloning, hence the use of 5' polyphosphatase treatment to reveal this population. TAP (tobacco acid pyrophosphatase) is another enzyme that removes 5' triphosphate but it also removes 5' CAP structures- we used this to look for piRNA precursor sequences in C. elegans that have a 5' cap.