NCBI GEO Submission of human whole blood transcriptomes in response to a high-fat meal

Modern humans spend most of their time having eaten recently. The purpose of the current project is to understand how the blood, which contains immune cells, responds in the hours after eating a meal that is moderately high in fat. We used a sequencing method to observe the expression of all the genes in blood cells in five participants who were each fed a high fat meal on three separate days. The results are reported in the manuscript, “Temporal changes in postprandial blood transcriptomes reveal subject-specific pattern of expression of innate immunity genes after a high-fat meal." Overall design: We used a sequencing method to observe the expression of all the genes in blood cells in five participants who were each fed a high fat meal on three separate days, resulting in 45 whole blood transcriptomes. For each sample, 3 mL of venous whole blood was drawn into a Tempus Blood RNA tube, shaken vigorously, and then frozen at -80°C until use. Total RNA was purified with the Tempus Spin RNA Isolation Kit with minor modifications to the manufacturer’s protocol. To remove residual genomic DNA, RNA samples were treated on-column with RNase-Free DNase per manufacturer’s instructions. RNA quantity, quality, and integrity were assessed with NanoDrop 1000 and 2100 Bioanalyzer. All isolated RNA had A260/A280 ratios greater than 2 and RNA integrity numbers higher than 7.3. RNA-Seq libraries were constructed at the DNA Technologies and Expression Core at the University of California, Davis, using the Ovation Human Blood RNA-Seq Library System (NuGEN Technologies). Sequencing was performed in a 2x100bp format with 45 samples multiplexed on 3 lanes on an Illumina HiSeq 4000. Analysis of the data is reported in the manuscript, “Temporal changes in postprandial blood transcriptomes reveal subject-specific pattern of expression of innate immunity genes after a high-fat meal.”