ez dna methylation kit  (Zymo Research)

Journal: bioRxiv

Article Title: Integration of Multiomic and Multi-phenotypic Data Identifies Biological Pathways Associated with Physical Fitness

doi: 10.1101/2025.09.24.677381

Figure Lengend Snippet: A. ACFT consists of six individual events (maximum deadlift (MDL), standing power throw (SPT), hand release pushups (HRP), sprint drag carry (SDC), leg tuck or plank (LTK), and two-mile run (2MR)) that are scored from 0 to 100 and summed into a total score, up to a maximum 600 points. B. Schematic representation of phenotypic measures including physiological, cognitive, behavior, hematology, and sleep. The exercise event was repeated weekly over five weeks along with random assignments to perform the exercise at three controlled time periods (morning, mid-day, late afternoon). Each exercise event consisted of a 20-minute strenuous cycling exercise at 70% VO2max with four blood draws, one at baseline, just prior to the bike exercise, and three draws approximately <5, 10, and 30 minutes after completion of the bike exercise. The types of molecular measurements (N=50,011) assayed from the drawn blood included DNA methylation of PBMCs, whole blood transcriptomics untargeted metabolomics and proteomics, and targeted biomarkers, including a cytokine panel. C . Cumulative distribution function (CDF) of the ACFT total points for the Spring 2021 USMA West Point male total population (thick black line, N=2,414); the study cohort of 65 male Cadets is also shown in the same plot (grey line, N=65). The vertical red dashed line is at the threshold value of 540, which is considered a differentiating point of physical fitness by the US Army. The horizontal black dashed line is at 0.5 or the median of the CDF. The male study cohort scored significantly higher than the larger USMA population (p=0.004, Kolmogorov-Smirnov test) D. Distribution of ACFT total points in the male study cohort, along with a vertical red dashed line at the 540 ACFT score cut-off; 60% of the male participants had scores above 540. E. Heatmap of the male cohort (rows) and their individual points for the six ACFT athletic events in the vertical columns (MDL, SPT, HRP, SDC, LTK, 2MR), where the darker grays indicate higher scores, and purple indicates a maximum score for that activity. F. CDF of the ACFT total points for Spring 2021 USMA West Point female total population (thick black line, N=493); the study cohort of female cadets is shown as the grey line (N=14). The vertical red dashed line is at the threshold value of 540 and the horizontal black dashed line is the median of the CDF. The female study cohort scored significantly higher than the larger USMA population (p=0.003, Kolmogorov-Smirnov test). G. Distribution of ACFT total points in the female study cohort, along with a vertical red dashed line at the 540 ACFT score cut-off. H. Heatmap of the female cohort (rows) and their individual points for the six ACFT athletic events in the vertical columns (MDL, SPT, HRP, SDC, LTK, 2MR), where the darker grays indicate higher scores, and purple indicates a maximum score for that activity.

Article Snippet: During the bisulfite conversion step, the gDNA was deaminated utilizing the EZ DNA Methylation kit (Cat# D5003, Zymo Research) in accordance with Illumina’s preferred deamination protocol.

Techniques: DNA Methylation Assay, Activity Assay