GSM5149474: MOLM14 Parental R3; Homo sapiens; RNA-Seq
RNA was extracted using Qiagen miRNAeasy kit RNA was run on a Bioanalyzer to assess quality. Libraries were prepared from total RNA using the TruSeq Stranded mRNA Kit (Illumina). Briefly, poly(A)+ was extracted using oligo-dT-coated beads, then fragmented by treatment with divalent cations and heat. Double stranded cDNA was then generated using random hexamer primers. Second strand synthesis was done with dUTP replacing dTTP to generate strand-specificity. The cDNAs were ligated to sequencing adapters and amplified using limited rounds of polymerase chain reaction. The libraries were quantified using real time PCR on an ABI StepOnePlus Real Time PCR Workstation (Thermo) using a Kapa Library Quantification kit (Roche).  Libraries were sequenced on a NovaSeq 6000 (Ilumina). Fastq files were assembled using Bcl2Fastq (Ilumina).