GSM8744884: Nfat5-KD MEF unstim RNA-seq (replicate 3); Mus musculus; RNA-Seq
Cells were washed once with PBS and total RNA was isolated with TriZol following manufacturer's recommendations. RNA concentration was quantified by ND-1000 spectrophotometer (Nanodrop). DNA libraries were prepared from 1 ug of total RNA using KAPA mRNA Hyperprep Kit (Roche). PolyA+ mRNA was enriched and reverse transcribed to double stranded cDNA. The cDNA was end repaired, adenylated, and ligated with KAPA Unique Dual-Indexed adapters (Roche). Libraries were amplified by PCR for 8 cyclces and quantified by dsDNA Qubit (Thermo) before pooling and sequencing.