GSM7728623: Mouse_2996-2_heart_AAV_miR-29a; Mus musculus; RNA-Seq
Total RNA was isolated from TRIzol according to the manufacturer's instructions (Thermo Fisher) and dissolved in nuclease-free water and quantified on a Nanodrop Spectrophotometer. RNA was DNase-treated using a DNA-free kit (Thermo Fisher). RNA quality assessment and sequencing were performed by the Iowa Institute of Human Genetics, Genomics Division, at the University of Iowa. RNA quality and concentrations were determined with the Agilent 2100 Bioanalyzer, and DropSense or Little Lunatic (Unchained Labs, Pleasanton, CA) reads. The RNA integrity number (RIN) values for samples were >7.5. Illumina TruSeq® Stranded mRNA Library Prep kits (Illumina) were used to prepare sequencing libraries from mRNA, followed by reverse transcription to cDNA, fragment purification and ligation to indexed (barcoded) adaptors. Indexed libraries were pooled for multiplex sequencing and run on an SP flow cell on the Illumina NovaSeq 6000 sequencer for 100 cycles with 50-base paired-end reads.