RNA was isolated using TRIzol (Invitrogen), followed by RNA clean-up using the QIAGEN Rneasy mini kit with on-column DNAse treatment (Venlo, the Netherlands). RNA quality and concentration were measured using an Agilent 2100 Bioanalyzer (Santa Clara, CA, USA) and Nanodrop ND-1000 (Thermo Fisher Scientific, Waltham, MA, USA), respectively.
Label
biotin
Label protocol
Synthesis of cDNA, cRNA amplification, and hybridization of cRNA onto the Illumina Human-6 v2.0 Expression BeadChips (San Diego, CA, USA) were performed as per manufacturer's instructions.
Hybridization protocol
Synthesis of cDNA, cRNA amplification, and hybridization of cRNA onto the Illumina Human-6 v2.0 Expression BeadChips (San Diego, CA, USA) were performed as per manufacturer's instructions.
Scan protocol
The slides were scanned on a BeadArray Reader (Illumina, Inc.).
Description
Two normal bone samples from different donors were purchased from Capital Biosciences (Gaithersburg, MD, USA).
Data processing
The data was extracted and quality controlled using the Gene Expression module v3.1.7 of Illumina´s BeadStudio software (v3.1.0.0). Variance-stabilizing transformation (Lin et al., 2008) and quantile normalization on the probe level was carried out using the statistical package R and the Bioconductor package lumi (Du et al., 2008; Gentleman et al., 2004).
