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1st Student's Major

Chemistry and Geology

1st Student's College

Science, Engineering and Technology

Students' Professional Biography

Cassidy Punt began his pursuit of a Bachelor of Science Degree in Biochemistry from Minnesota State University, Mankato in the spring of 2002. His research began in the spring of 2005 with the help of Elizabeth Smalley, a Minnesota State University, Mankato alumna (2005) of the Department of Chemistry and Geology. In the summer of 2005, he was called to active duty with the Minnesota Army National Guard to serve in Iraq. Upon returning in the summer of 2007, he continued in his research and education within the Biochemistry program at Minnesota State University, Mankato. His research was completed in the spring of 2008 and was presented as a poster at the 10th Annual Undergraduate Research Conference at Minnesota State University, Mankato. Following graduation in the fall of 2008, Cassidy plans to earn a Ph.D. in human or medical genetics in order to pursue a career of his own in teaching and research.

Mentor's Name

Theresa Salerno

Mentor's Email Address

theresa.salerno@mnsu.edu

Mentor's Department

Chemistry and Geology

Mentor's College

Science, Engineering and Technology

Abstract

Two novel single nucleotide polymorphisms (SNPs) were discovered within the coding region of the NADH dehydrogenase subunit 2 gene of mitochondrial DNA (mtDNA). mtDNA is of particular importance in forensic analysis as well as in the study of the origin and dispersal of humans. Two segments of the coding region of human mtDNA, as well as the hyper-variable region 2 (HV 2) were selected and sequenced in order to determine if any previously unknown SNPs were present in our test subjects. Target regions were designed to include known SNPs; appropriate primers were developed using the OLIGO 6 Primer Analysis Software. The DNA was isolated using the QIAGEN® Generation Capture Column DNA Extraction kit and target regions were amplified via polymerase chain reaction (PCR). Once the fragment sizes were verified using acrylamide gel electrophoresis, the template DNA was prepared for sequencing by PCR using forward primers and IRDyeTM labeled dideoxy-terminators. Sequencing PCR products were then purified to remove excess primers and nucleotides and sequenced with the LICOR NEN model 4300 slab-gel DNA sequencer. The SNP analyses developed in this research have been implemented into the Minnesota State University, Mankato biochemistry laboratory curriculum. The two novel SNPs identified were: T644G and T4733G inside the NADH dehydrogenase subunit 2 gene. Each of these occurred in only one of our test subjects. No novel SNPs were found in the HV 2 region. However, sequencing was successful for only one of DNA test samples. Seven previously known SNPs were also found: G3010A and C3116T contained in the 16S rRNA coding region; T4216C contained in the NADH dehydrogenase 1 subunit gene; C4312T contained in the isoleucine tRNA gene; G4491A, T4586C, and T4688C contained in the NADH dehydrogenase 2 subunit gene.

Creative Commons License

Creative Commons Attribution-Noncommercial 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial 4.0 License

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Genetics Commons

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