Rneasy mini kit with rnase free dnase treatment

Genomic DNA was extracted from peripheral blood using the QIAamp DNA blood Mini Kit (Qiagen, Crawley, UK) and total RNA was harvested using the RNeasy^® Mini Kit with RNase-free DNase treatment (Qiagen) and analysed as previously published (Smith et al., 2015 (link)).

Total RNA was harvested from MDM using the RNeasy Mini Kit with RNase-free DNase treatment (Qiagen, Hilden, Germany), in accordance with the manufacturer's instructions. Optical density readings were determined for OD₂₆₀/OD₂₈₀ and OD₂₆₀/OD₂₃₀ using a NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, Hemel Hempstead, UK) to assess protein and solvent contamination, respectively. RNA integrity was analysed by assessing ribosomal RNA band 28S/18S ratios using high-resolution electrophoresis on an Agilent Bioanalyser (Agilent Technologies, Inc, Berkshire, UK) and attained an integrity score of > 8. Total RNA (500 ng) was amplified using the Illumina TotalPrep-96 RNA Amplification kit (Life Technologies Ltd, Paisley, UK), normalized to 150 ng/μl, and 750 ng was hybridized to Illumina Human-WG6 v3.0 Expression BeadChips (Illumina, Essex, UK) for 16 hr at 58°. Bead arrays were stained with streptavidin-Cy3 (GE Healthcare, Amersham, UK) and scanned using the Bead array reader and processed with genomestudio® data analysis software (Illumina). Data were smoothed with cubic spline normalization.