Term of Award

Summer 2021

Degree Name

Master of Science in Biology (M.S.)

Document Type and Release Option

Thesis (restricted to Georgia Southern)

Copyright Statement / License for Reuse

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.


Department of Biology

Committee Chair

Aaron Schrey

Committee Member 1

Andrea Liebl

Committee Member 2

Alexander Collier


Epigenetic mechanisms can alter gene expression without altering the underlying DNA sequence. DNA methylation is the most well-studied epigenetic mechanism in an ecological setting. Ecological epigenetics has provided insight into invasive species, behavioral and physiological phenotype expression, and developmental changes based on environmental factors. The cooperative breeding Chestnut-crowned babbler (Pomatostomus ruficeps) is an excellent model to investigate ecological epigenetics. A previous study of epigenetic factors relating to their behavioral phenotypes found variation in DNA methylation based on behavior and developmental stage. Additional studies of DNA methylation leveraging techniques that offer greater precision on the methylation status of individual cytosines is needed. Here, I optimized a protocol using Enzymatic-Methyl Sequencing (EM-seq). I screened four birds at three different developmental stages, creating a total of 12 libraries for sequencing on the Illumina MiSeq platform. EM-seq resulted in an average of 763,521.92 non-duplicate reads for each sample, an average GC content of 25.9, and a total of 495,818,189 bases in a DeNovo assembly. Now that this protocol has proved successful, further studies are possible. The future directions include a whole-genome survey of differently methylated regions, and a targeted approach assaying the methylation status of cytosines located in specific promoter regions of genes of interest.

OCLC Number


Research Data and Supplementary Material