title: GSM8440921: MLE-12_Mir9LOF_G4_CaT; Mus musculus; OTHER
library_construction_protocol: Cells were transfected with Anti-hsa-miR-Mir9-5p (Ambion, #17000 were transfected at 60 nM final concentrations using Lipofectamine 2000 (Invitrogen) following the manufacturer's instructions and harvested 48 h later for further analysis. TruSeq ChIP Library Preparation Kit (Illumina) with the following modifications. Instead of gel-based size selection before the final PCR step, libraries were size selected by SPRI-bead-based approach after the final PCR with 18 cycles. In detail, samples were 1stcleaned up by 1x bead: DNA ratio to eliminate residuals from PCR reaction, followed by a 2-sided-bead cleanup step with an initial 0.6x bead:DNA ratio to exclude larger fragments. The supernatant was transferred to a new tube and incubated with additional beads in 0.2x bead:DNA ratio to eliminate smaller fragments, like adapter and primer dimers. Bound DNA samples were washed with 80% ethanol, dried, and resuspended in TE buffer. Library integrity was verified with LabChip Gx Touch (Perkin Elmer).