RNA-seq of homo sapiens: foreskin bulk sequencing
Tissue samples were disrupted and homogenized using a Tissuelyzer (Qiagen) and total RNA isolated using the RNeasy Kit (Qiagen) according to manufacturers instructions. RNA was subjected to quality control with Agilent Bioanalyzer (Agilent). To construct libraries suitable for Illumina sequencing, the Illumina stranded mRNA prep ligation sample preparation protocol was used with starting concentration of 200 ng total RNA. The protocol includes mRNA isolation, cDNA synthesis, ligation of adapters and amplification of indexed libraries. The yield and quality of the amplified libraries were analysed using Qubit by (Thermo Fisher) and the Agilent Tapestation (Agilent). The indexed cDNA libraries were normalized and combined, and the pools were sequenced on the Illumina Novaseq 6000 S4 flowcell for a 150 cycles paired-end sequencing run generating 150 bp paired-end reads. A single tissue sample was obtained for each participant included in the study from which transcriptome was derived.