title: GSM8806683: MDA-MB-231/Res antiIGF2BP2 Replicate 1; Homo sapiens; RIP-Seq
library_construction_protocol: For the RIP-seq experiment, MDA-MB-231/Sens and MDA-MB-231/Res cells were harvested in RIP lysis buffer. The equivalent lysates were firstly incubated with 5μg anti-IGF2BP2 mAb or 5μg rabbit IgG overnight, and then captured by Protein A/G agarose for additional 4 hours at 4°C. Next, the rinsed agaroses were probed to TRIzol RNA extraction. The purified RNA was qualified and quantified. Equivalent amount of RNA was mixed with spike-in (Invitrogen #4456740) to generate RIP-IGF2BP2 libraries using VAHTS Universal V6 RNA-seq Library Prep Kit (Vazyme #NRM604-01), according to the manufacture instructions. The libraries were probed to sequencing. For bioinformatic analysis, the sequencing data were normalized to the signal of spike-in (ERCC mixture) using RUVSeq's method and the circRNA enrichment was calculated by comparing the reads numbers in IGF2BP2 groups (Sens and Res) to the IgG counterparts.
identifiers.namespace: GEO
identifiers.id: GSM8806683_r1