title: Illumina HiSeq 4000 paired end sequencing; Context-specific regulation of monocyte surface IL7R expression and soluble receptor secretion by a common autoimmune risk allele
design: Context-specific regulation of monocyte surface IL7R expression and soluble receptor secretion by a common autoimmune risk allele
library_construction_protocol: Peripheral blood was obtained from genotyped individuals recruited via the Oxford biobank (www.oxfordbiobank.org.uk) with full ethical approval (REC: 06/Q1605/55) and written informed consent. Genotype was blinded at the time of study and only revealed at the end of recruitment. Peripheral blood and synovial fluid samples from were recruited with informed consent from patients with inflammatory arthritis attending the Oxford University Hospitals NHS Foundation Trust (Ethics reference number 06/Q1606/139).  The AllPrep DNA/RNA/miRNA kit (Qiagen) was used for RNA extraction. Cells were spun down and re-suspended in 350μl of RLTplus buffer and transferred to 2ml tubes. Samples were then stored at -80°C for batched RNA extraction. Homogenization of the sample was carried out using the QIAshredder (Qiagen). DNase I was used during the extraction protocol to minimise DNA contamination. RNA was eluted into 35μl of RNasefree water. The RNA amount was quantified by qubit and the RNA samples stored at - 80°C for storage until ready for sequencing.  RNA underwent quality control testing using a bioanalyser (RNA 6000 Nano kit, Agilent) followed by cDNA library preparation.
attributes.tag: Experimental Factor: stimulus
attributes.value: none