Biological clock measures: Assessing the association between the circadian and epigenetic clock as predictors of migration phenology and biological aging in wildlife

This study will investigate the relationship between clock genes and migration, as well as epigenetics and aging. We will study repeats in a poly Q residue of the Clock gene and repeats in the five prime UTR of the Adcyap1 gene to study migratory phenology. Epigenetic age studies will be done by quantitating methylation levels at different ages in relevant CpG residues, and by assaying telomere length, to construct age determining models. DNA will be extracted from blood with commercial kits. PCR will be performed on clock genes with reported primers modified to the study species. PCR products will be purified by Exonuclease I and Shrimp Alkaline Phosphatase digestion and sequenced with the ABI 3130 Genetic Analyzer. Methylation analysis will be done for 8 CpGs via bisulphite sequencing. An age estimating model will be constructed from strong correlating sites. Absolute telomere length will be measured by qPCR. These values will be used to model age.