title: RNA-seq of Human sapiens
design: We referred to the Smart-seq2 protocol (Picelli, S. et al. 2014) for all RNA-seq experiments Single cells were washed twice with 1XPBS before being placed in lysis buffer. RNA was isolated from single cells and converted into cDNA. Library construction was performed following the Smart-seq2 protocol, and sequenced reads that contained polyA, low quality, and adapters were pre-filtered before mapping. cDNA was sheared into 100- to 150-bp short fragments according to the manufacturers instructions. Libraries were pooled and sequenced on Illumina HiSeq2500 sequencers. Data normalization was carried out by transforming mapped transcript reads to fragments per kilobase of transcript per million mapped reads (RPKM). Genes with fragments per kilobase of transcript per million mapped reads (FPKM) >0.5 were retained for analysis.