title: GSM8794014: SRSF10 Input sample, primary rat DRG neurons, bio rep 4; Rattus norvegicus; RIP-Seq
library_name: GSM8794014
library_construction_protocol: RNA Immunoprecipitation was performed according to the EZ- Magna RIP RNA-Binding Protein Immunoprecipitation (Millipore Sigma) protocol. In brief, on Day 5 of primary DRG neuron cell culture, cells were washed with ice-cold PBS and lysed at -80 °C overnight in Complete Lysis Buffer. Lysates were spun down to pellet cellular debris and the supernatant added to Protein A/G Beads conjugated with 5μg of primary anti-SFPQ (Abcam, #ab38148), anti-SRSF10 (MBL, #50304712), or anti-rabbit IgG (EZ-Magna Kit). Lysate-bead mixtures were incubated at 4 °C overnight with end-to-end rotation. Protein-RNA complexes were eluted from beads in a Proteinase K Buffer for 30 mins at 55 °C. RNA was further purified using phenol:chloroform:isoamyl alcohol procedure followed by absolute ethanol and salt RNA precipitation. Libraries were prepared with Takara SmartSeq v4 full length cDNA synthesis using oligo dT priming & 5' template switching enzyme.