Flashtag biotin hsr labelling kit

Global miRNA expression was examined using microarrays in a subset of participants at all timepoints (N=32; 8 participants, 4 timepoints). Samples were selected from the 8 participants with the highest RNA quantity and purity across all timepoints (Concentration: 25-92.6 µg/mL; 260/280: 1.9-2.1 nm). 100 ng RNA were labelled using the Flashtag Biotin HSR labelling kit (ThermoFisher Scientific, UK, cat no. 901910).
Hybridization cocktails were prepared using the GeneChip hybridization kit (ThermoFisher Scientific, UK, cat no. 900720). Biotin labelled samples were hybridized overnight onto GeneChip miRNA 4.0 arrays (ThermoFisher Scientific, UK, cat no. 902413) according to manufacturer's instructions. The arrays were washed and stained using the GeneChip wash and stain kit (ThermoFisher, UK, cat no. 902413) and the Affymetrix fluidics station 450, as per manufacturer's instructions. GeneChips were scanned by the Affymetrix scanner 3000 7G. Microarray signal intensities were normalized using the variance stabilization normalization 2 (VSN2) algorithm and probe level summarization conducted using robust multi-chip analysis (RMA) (31) (link). Microarray data has been deposited to the Gene Expression Omnibus (GEO) with the accession number GSE180625.