title: NextSeq 550 paired end sequencing; RNA sequencing of unmanipulated and saline-injected skin in healthy old adults before and after oral vitamin D3 supplementation
design: RNA sequencing of unmanipulated and saline-injected skin in healthy old adults before and after oral vitamin D3 supplementation
library_construction_protocol: 3 mm punch skin biopsies were taken and collected into RNAlater (Qiagen) for RNA extraction. Frozen tissue was homogenized using a Precellys Evolution homogenizer (Bertin Instruments), and total RNA was extracted with the RNeasy Mini Kit (Qiagen, Hilden, Germany). RNA was subjected to DNase treatment to remove contaminating genomic DNA using a TURBO DNA-free kit (Ambion, Life Technologies).  cDNA libraries were generated using the Kapa mRNA Hyperprep kit (Roche).