GSM8332733: PHGDH-1; Homo sapiens; RIP-Seq
RNA was precipitated by the addition of 3 µL of glycogen (5 mg/mL, Invitrogen™, AM9510), 50% of 2-propanol (Sigma-Aldrich, I9516-500ML), and 10% of 3 M sodium acetate, pH 5.5 (Invitrogen™, AM9740) with an incubation at -80℃ overnight. The RNA was then pelleted by centrifugation at 14k rpm for 30 minutes at 4℃, washed with 1 mL of 75% ethanol (Sigma-Aldrich, 493546), and air-dried. The RNA was then suspended in 20 µL of UltraPure™ DNase/RNase-Free distilled water (Invitrogen™, 10977015). A library was prepared by cDNA synthesis, amplification, fragmentation, and adaptor ligation using NEBNext® Low Input RNA Library Prep Kit (NEB, E6420) and sequenced 150 base pairs from each end on an Illumina MiniSeq.