Out of thousands of known millipede species, only five sequenced genomes (in four of sixteen orders) are publicly available. No whole genomes and limited genetic information are available for incredibly diverse families such as Xystodesmidae. The purpose of this project (our research goal) is to sequence the whole genome of the millipede Cherokia georgiana. A de novo sequence of the complete genome of a North American species will facilitate future research in understanding gene expression under a variety of conditions. Many interesting biological processes in millipedes are poorly described, such as the production of a defensive hydrogen cyanide secretion found in the Polydesmida. While genes in this pathway have been identified, it is unclear how they differ between polydesmid families. Another research avenue this will facilitate is understanding how this cyanogenic pathway is regulated under stressed conditions. This may also aid in understanding this species’ importance in phylogeny and its relationship to other North American millipede species. Here, we present our research strategy for de novo sequencing using the next-generation sequencing platform Oxford Nanopore MinION. For this, DNA extraction kits will be used to extract high-molecular weight (HMW) DNA to be used in sequencing. We will evaluate NGS library quality using the Agilent TapeStation 4200 automated electrophoresis system. Long read sequencing data will be collected using multiple flow cells to enhance read-depth and accuracy. Genome assembly will be conducted using available millipede genome data, and genomes of related species. In future experiments, we will conduct RNA-seq analysis to identify actively transcribed genes.


GCSU Department of Biological and Environmental Sciences

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