PBMCs were isolated, pellets were stored at -80 °C before mRNA was extracted.
Extracted molecule
total RNA
Extraction protocol
mRNA was extracted using RNeasy Mini Kit (Qiagen) acording to the protocol provided. RNA quantity was measured using NanoDrop-1000 (NanoDrop Technologies, Inc., Delaware, USA), while RNA quality was checked with Aglient 2100 Bioanalyzer (Agilent Technologies, Inc., California, USA).
Label
biotin
Label protocol
Illumina Total Prep RNA Amplification Kit (Illumina Inc., California, USA)
Hybridization protocol
Illumina HumanHT-12 v4 Expression BeadChip (Illumina Inc., California, USA)
Scan protocol
Illumina HiScan System (Illumina Inc., California, USA)
Data processing
The data was normilised using the Illumina GenomeStudio software, version 1.7.0. Genome Studio Illumina was used to calculate and report a detection p-value, which represents the confidence that a given transcript is expressed above the background. A gene was defined as expressed when relevant probes with a p-value below 0.01 were found in more than five samples. After hybridization and scanning, a manual quality control was performed investigating density plots and hierarchical clustering of raw probe densities. One probe per gene (max IQR) was selected for further analysis.
Intake of protein modulate immune function, POMC processing and IGF signaling gene expression pathways in peripheral mononuclear cells in home-dwelling old subjects - a transcriptomic approach
