title: GSM1152971: Immunoprecipitated mRNAs with anti-IGF2BP3 antibody; Homo sapiens; OTHER
library_construction_protocol: Fibronectin-stimulated S2-013 cells were lysed in NP2 buffer containing 50 mM Tris (pH 7.4), 150 mM NaCl, 1 mM MgCl2, 0.5% NP-40 and protease and RNAse inhibitors. RIP protocol: Cell lysates were immunoprecipitated with anti-IGF2BP3 antibody or with normal rabbit IgG and Dynabeads Protein G (Dynal, Oslo, Norway) for 2 h at 4ºC. Beads were pelleted on a magnetic rack (Dynal). The immunoprecipitation products were treated with proteinase K, extracted with phenol-chloroform, and precipitated with ethanol. Purified RNAs were subsequently treated with DNase I (Promega, Madison, WI) and extracted using the RNeasy kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions. Finaly, RNAs were fragmented and purified using an Illumina TruSeq RNA Sample Prep Kit (San Diego, CA) according to the manufacturer’s instructions. seq protocol: The IGF2BP3-associated mRNAs were identified using a next generation sequencer, Illumina HiSeq 2000 (Hokkaido System Science, Sapporo, Japan).
attributes.tag: GEO Accession
attributes.value: GSM1152971