GSM5739959: MDAMB231_Static_rep3; Homo sapiens; RNA-Seq
GSM5739959
Cells were lysed directly on microfluidic channels in RLT lysis buffer and total RNA was isolated (RNeasy Micro kit, Qiagen), according to the manufacturer's instructions. RNA was measured using a Qubit RNA high sensitivity assay and an Agilent Bioanalyzer total RNA 6000 Pico chip. A KAPA mRNA HyperPrep protocol for Illumina platforms was used for poly(A) mRNA capture and construction of reverse-stranded mRNA-Seq libraries. Briefly, mRNA was captured using oligo-dT beads, fragmented with heat and magnesium, and cDNA synthesized according to manufacturer guidelines. Libraries were ligated to KAPA adapter sequences and amplified. cDNA was quality checked on an Agilent Bioanalyzer high sensitivity DNA assay chip. Equimolar amounts of each sequencing library were pooled and purified with magnetic KAPA Pure Beads. Pool was sequenced on the Illumina NextSeq 550 using the NextSeq500/550 High Output kit v2.5 (2x75 paired end cycles).