GSM8666380: eCLIP-seq, LS174T, mouse PXR, IP, replicate 2; Homo sapiens; RIP-Seq
RNA fragments were extracted using acid phenol:chloroform:isoamyl alcohol (25:24:1, pH 6.6) (Invitrogen) and cleaned up RNA Clean & Concentrator-5 columns (Zymo). After reverse transcription, qPCR was performed to determine the Ct value of the cDNA with Power SYBR Green Master Mix (Applied Biosystems). DNA fragments were amplified with primers containing Illumina adapter sequences using Q5 High-Fidelity 2× Master Mix (NEB) for 9 to 18 cycles based on the Ct values, and PCR products were purified with Agencourt AMPureXP beads (Beckman Coulter). The DNA fragments ranging from 175 - 300 bp were selected via agarose gel electrophoresis. Fragments was excised and extracted using the MinElute Gel Extraction Kit (Qiagen) and libraries were validated using the 1× High Sensitivity DNA Kit (Invitrogen).