GSM1533654: Flag-Siwi bound piRNAs from Vasa siRNA treated BmN4; Bombyx mori; RIP-Seq
Siwi, BmAgo3, and Flag-Siwi complexes were immunopurified from BmN4 cells (untreated or treated with indicated siRNAs) using anti-Siwi, anti-BmAgo3, and anti-Flag antibody bound to Dynabeads protein G (Invitrogen). The beads were washed three times with binding buffer [30 mM HEPES (pH 7.4), 150 mM potassium acetate, 5 mM magnesium acetate, 5 mM DTT, 0.1 % NP-40, 2 mg/mL pepstatin, 2 mg/mL leupeptin and 0.5% aprotinin] containing 500mM sodium chloride and twice with binding buffer. Total RNAs were eluted from the beads using phenol–chloroform and precipitated using ethanol. small RNA-seq libraries were prepared using NEBNext Small RNA Library Sample Prep Set (New England BioLabs (NEB), E7330).
GEO Accession
GSM1533654