GSM7963890
Tissues were lysed and homogenized in TRIzol Reagent (ThermoFisher) using a Pellet Pestle Motor (Kimbal), then RNA was isolated using the miRNeasy Micro kit (Qiagen), according to manufacturers' protocols, including the optional DNase digest step.  RNA concentration and quality were assessed using the Nanodrop 2000/Nanodrop 8000 spectrophotometer (Thermo Scientific) and the RNA 6000 Nano and Pico Assay (Agilent Technologies). Libraries were constructed using the KAPA mRNA HyperPrep Kit (Roche Sequencing and Life Science), according to the manufacturer's protocol.  Briefly, the protocol entails isolation of polyA containing mRNA using oligo-dT magnetic beads, RNA fragmentation, first and second strand cDNA synthesis, ligation of Illumina-specific adapters containing a unique barcode sequence for each library, and PCR amplification.  The quality and concentration of the libraries were assessed using the D5000 ScreenTape (Agilent Technologies) and Qubit dsDNA HS Assay (ThermoFisher), respectively, according to the manufacturer's instructions. Libraries were sequenced 150 bp paired-end on an Illumina NovaSeq 6000 using the S4 Reagent Kit v1.5.