RNA-sequencing transcriptome profiling of normal human keratinocytes differentiation

In this study, we used a RNA-sequencing (RNA-seq) approach to analyze the whole transcriptomes of human primary Keratinocytes (KC) at different differentiation stages. An average of 72.56 million reads were collected for each sample: 87.68% of the sequences could mapped to the human genome, and 66.70% of sequence mapped to known human genes. A total of 17,446 ± 220 genes were expressed during the course of differentiation. 1024 transcription factors (TF) and genes encoding TF binding proteins were detected during the course of differentiation were expressed during the course of KC differentiation. Overall design: Foreskin normal human keratinocytes were differentiated in vitro by increasing Ca2+ concentration in culture media from 0.06mM to 1.3 mM. Undifferentiated cells and cells under differentiation for 24 hrs, 48hrs, 72 hours, 96 hours and 120 hours were harvested for total RNA extraction. RNA sequencing libraries were made using eukaryotic long non-coding RNA sequencing library construction protocol. RNA libraries were deep sequenced by 100bp paired-reads on Illumina His-seq 2000.