title: GSM8146057: NHEK_CTLi_1; Homo sapiens; RNA-Seq
library_construction_protocol: Primary neonatal human epidermal keratinocytes were cultured in KGM-Gold media (Lonza). Cells were infected with either CASZ1 shRNA or control shRNA lentiviral particles, and selected in the presence of 0.4ug/ml puromycine. To induce epidermal differentiation, growth media was supplemented with 1.4mM CaCl2 and cells were grown for six days. Cells were then dissociated by warm 0.25% Trypsin for 10 min at 37°C. Cells were then centrifuged at 200 g for 4 min at 4°C, washed twice with 1x PBS and incubated in 350 µl Trizol for 10 min at room temp. Total RNA was purified using TriRNA Pure Kit (Geneaid) according to manufacturer's instructions. of total RNA per sample was used. Enrichment for poly(A) mRNA was done using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs). The RNA-seq libraries were then constructed following standard protocol for the NEBNext Ultra II RNA Library Prep Kit for Illumina (New England Biolabs).  The concentration and quality of the libraries were determined using Qubit (Invitrogen) and Bioanalyzer (Agilent). Constructed RNA-seq libraries were sequenced on the Illumina NextSeq 2000 platform, obtaining 100-nucleotide single reads.