title: Illumina NovaSeq 6000 paired end sequencing; RNA-seq of human leukemia cell lines, MHH-CALL-4 and MUTZ-5, treated either with Ruxolitinib, FX1, or their combination.
design: RNA-seq of human leukemia cell lines, MHH-CALL-4 and MUTZ-5, treated either with Ruxolitinib, FX1, or their combination.
library_construction_protocol: Cells treated with drugs for 24hr were collected Cells were grown is RPMI1640-10%FCS media.  Cells were treated with either ruxolitinib (JAK1/2 inhibitor), FX1(BCL6 inhibitor), their combination, or DMSO(control) for 24 hrs. Qiagen RNAeasy-plus mRNA enriched using oligo(dT) beads was fragmented randomly by adding fragmentation buffer, then the cDNA is synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. Second, after a series of terminal repair, A ligation and sequencing adaptor ligation, the double-stranded cDNA library is completed through size selection and PCR enrichment.