GSM2498543: WT Pum1 iClip rep2; Mus musculus; RIP-Seq
0.5g neonatal mouse brain tissues were homogenized in PBS and crosslinked 400mj/cm2 at 254nm twice. 10 -7 dilution of RNase1 was used to trim RNA prior to IP with anti-Pum1/2 antibody. The other steps were performed as described in (Konig et al. 2011, Journal of visualized experiments: JoVE). The PCR cycle numbers were optimized for each sample to generate comparable amount of DNA. Please refer to Supplemental Figure S6 for the PCR cycle numbers for each individual sample. RT primers are WT_P1_clip /5Phos/NNGCCANNNAGATCGGAAGAGCGTCGTGgatcCTGAACCGC; P1KO_P1_clip /5Phos/NNCGCCNNNAGATCGGAAGAGCGTCGTGgatcCTGAACCGC; WT_P2_clip /5Phos/NNAACCNNNAGATCGGAAGAGCGTCGTGgatcCTGAACCGC; P2KO_P2_clip /5Phos/NNATTGNNNAGATCGGAAGAGCGTCGTGgatcCTGAACCGC