4pm natural light + house light

Our shortened, final barcode (COI subunit)

DISCUSSION

We successfully completed our original goal, which was to obtain the CO1 subunit and test it as a successful barcode. We checked our result with the traditional and more commonly known barcode. Our shorter sequence matched with 100% accuracy. In the future, if other species could be barcoded in the same shorter fashion, the taxonomic process would be shorter, more cost-efficient, and easier for non-experts to do.

Additionally, we set out to find out what skill level was required to test a known barcode. We concluded that with cursory biology knowledge (knowing how to run a PCR and having access to some biotechnologies), one can perform this procedure with relative ease.

In the case of an outbreak of the ALB or other invasive species, a barcode allows quick and correct identification. Barcoding is a modern and effective taxonomic method. In the future, scientists hope to barcode all species.

LITERATURE CITED

1. “Anoplophora glabripennis cytochrome oxidase subunit I (COI) gene, partial cds; Mitochondrial.” National Institute for Biotechnology Information. USA.gov, n.d. Web. 20 Feb. 2013. <http://www.ncbi.nlm.nih.gov/nuccore/111414684>. 2. Anoplophora Glabripennis mitochondrion, complete genome. National Center for Biotechnology Information. USA.gov, n.d. Web. 20 Feb. 2013. <http://www.ncbi.nlm.nih.gov/nuccore/ NC_008221.1>. 3. “Asian Longhorned Beetle.” ODNR Division of Forestry. N.p., n.d. Web. 13 Feb. 2013. <https://ohiodnr.com/DNN/health/asianlonghorned/ tabid/5197/Default.aspx>. 4. “Asian Longhorned Beetle Biology and Ecology.” USDA Forest Service. N.p., n.d. Web. 13 Feb. 2013. <http://nrs.fs.fed.us/disturbance/invasive_species/ alb/biology_ecology/>. 5.”DNA Sequencing Sample Submission Guidelines.” Genewiz. N.p., n.d. Web. 3 Mar. 2013. <http://www.genewiz.com/public/Sample-Submission-Guideline.aspx>. 6. “DNeasy Blood & Tissue Kit.” Qiagen. N.p., n.d. Web. 20 Feb. 2013. <http://www.qiagen.com/Search/ DNeasy-Blood-and-TissueKit#orderinginformation>. 7. “Protocol for Taq 2X Master Mix.” New England Biolabs. N.p., n.d. Web. 20 Feb. 2013. <https://www.neb.com/protocols/2012/09/13/protocol-for-taq-2x-master-mix-m0270>. 8. “Resuspending PCR Primers.” The Functional Genomics Research Stream. U of Texas at Austin, n.d. Web. 20 Feb. 2013. <http://fg.cns.utexas.edu/fg/protocol__resuspending_PCR_primers.html>. 9. “USB® ExoSAP-IT® PCR Product Cleanup Product Numbers 78200/01/02/05/50 Brief Protocol.” affymetrix.com. N.p., n.d. Web. 3 Mar. 2013. <http://media.affymetrix.com/support/ technical/usb/brief_proto/78200B.pdf>. 10. “Using DNA Barcodes to Identify and Classify Living Things: Introduction.” DNA Barcoding 101. Cold Spring Harbor Laboratory., n.d. Web. 13 Feb. 2013. <http://www.dnabarcoding101.org/introduction. html>. 11.”What is DNA Barcoding?” Barcode of Life. N.p., n.d. Web. 13 Feb. 2013. <http://www.barcodeoflife. org/content/about/what-dna-barcoding>.

ACKNOWLEDGEMENTS

We would like to thank our teacher Mr. Maxwell for helping us with the project as a whole and allotting us time in class to complete it. We would also like to thank also Dr. Morgan Thompson and Luke De of the Pingry Biology Department. We appreciate your guidance and help while planning our project, designing primers, sending in orders, and the overall project as a whole. We would also like to thank Paul Moore and anyone at the US Forest Service who helped provide the beetles for us. Lastly we would like to thank Pingry for funding our project and allowing us to buy the necessary materials.