Mediator Subunit 30 Directs Myc Oncogenesis [RNA-seq]

Mediator Subunit 30 Directs Myc Oncogenesis [RNA-seq] RNA seq in Mia PaCa-2 was performed with 0.5ug/ml doxycycline-induced teton shRNA or cDNA overexpression, or pLKRO.1 shRNA knockdown directly. RNAseq in GSC3565 was performed in pLKO.1 shRNA knockdown. All shRNAs were packaged in lentivirus and infected into cells. RNA was extracted by Trizol (Life Technologies) RNA concentration and quality was quantified by Qubit and Agilent 2100 Bioanalyzer system, respectively. Libraries were generated using NEBNext Ultra II RNA Kits (NEB) and sequenced on the NovaSeq 6000 with 100 bp paired-end reads.

中介体亚基30调控Myc致癌进程[RNA测序（RNA-seq）]：针对Mia PaCa-2细胞的RNA测序实验，采用0.5μg/ml多西环素诱导的teton型短发夹RNA（short hairpin RNA，shRNA）或互补DNA（complementary DNA，cDNA）过表达体系，或直接通过pLKRO.1型shRNA进行基因敲降；针对GSC3565细胞的RNA测序实验，采用pLKO.1型shRNA进行基因敲降。所有shRNA均以慢病毒（lentivirus）包装后感染至宿主细胞。总RNA采用Trizol试剂（Life Technologies）提取，RNA浓度与质量分别通过Qubit定量仪与Agilent 2100生物分析仪进行检测。测序文库采用NEBNext Ultra II RNA建库试剂盒（NEB）构建，随后在NovaSeq 6000测序平台上开展100bp双端测序读长的测序工作。