GSM6569730: U87D0R2; Homo sapiens; RNA-Seq
The genome DNA was extracted by using QIAamp DNA Mini Kit (QIAGEN) for each replicate. Total RNA was isolated from U251 cells using RNeasy Mini Kit (QIAGEN #74104) and was treated with DNase I (QIAGEN #79254). Two rounds of PCR were employed to prepare the next-generation-sequencing (NGS)-ready sgRNA libraries with the KAPA HiFi HotStart ReadyMix (Roche # KK2602). The RNA-seq library were constructed with TruSeq Stranded mRNA Library Prep kit (Illumina # 20020594).The eCLIP libraries were generated from U251 cells in biological duplicates with the kit (Eclipse Bioinnovations Inc, #ECEK-0001) according to the standard single-end eCLIP protocol.