Transcriptome Sequencing of Hair Follicle and Interfollicular Epidermis

RNA for transcriptome sequencing was extracted from the 20 pooled microdissected HF of the same cycle stage and the IFE collected from each dog. RNA extraction, library preparation and RNA-seq were performed as described in a previous study [17 (link),36 (link)]. In brief, total RNA was extracted using the RNeasy Micro Kit (Qiagen, Hombrechtikon, Switzerland) including proteinase K digestion according to the manufacturer’s protocol. Total RNA content was measured using the Qubit^® 2.0 Fluorometer (Invitrogen, Thermo Fisher, Basel, Switzerland) and RNA quality was assessed with a Bioanalyzer (Agilent 2100; Agilent Technologies, Basel, Switzerland). High quality RNA (RIN > 9) was reverse transcribed into cDNA with the SMART-Seq v4 Ultra Low Input RNA Kit (Takara, Saint-Germain-en-Laye, France) and libraries were constructed following the TruSeq^® Nano DNA Library Prep (Illumina, San Diego, CA, USA) protocol for RNA sequencing. Multiplexed total cDNA libraries were sequenced on two lanes using an Illumina HiSq3000 instrument with 100 bp single-end sequencing cycles. On average, thirty million reads were collected, converted into FASTQ file format and demultiplexed.
The data are available from the European Nucleotide Archive (ENA), study accession number PRJEB21761 and sample accessions SAMEA7016531-SAMEA7016551 [50 ].