Identification of SNPs in the Coding Region of Human mtDNA
Location
CSU Ballroom
Start Date
21-4-2008 1:00 PM
End Date
21-4-2008 3:00 PM
Student's Major
Chemistry and Geology
Student's College
Science, Engineering and Technology
Mentor's Name
Theresa Salerno
Mentor's Department
Chemistry and Geology
Mentor's College
Science, Engineering and Technology
Description
Four novel single nucleotide polymorphisms (SNPs) were discovered in separate coding regions 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 (HV2) 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 Gentra column kit and target regions were amplified via the 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 IRDyeâ„¢ labeled dideoxy-terminators. Sequencing-PCR products were then purified to remove primers and sequenced with the LICOR NEN model 4300 slab-gel DNA sequencer. The SNP analyses developed in this research were used in two biochemistry laboratory classes. Four novel SNPs identified were: C4198A, found in the NADH dehydrogenase subunit 1 gene; T4586C, T4644G, T4688C 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 HV2 region, however, sequencing was successful for only one of our test subjects. Two previously known SNPs were also found: G3010A and C3116T which are both contained in the 16S rRNA coding region.
Identification of SNPs in the Coding Region of Human mtDNA
CSU Ballroom
Four novel single nucleotide polymorphisms (SNPs) were discovered in separate coding regions 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 (HV2) 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 Gentra column kit and target regions were amplified via the 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 IRDyeâ„¢ labeled dideoxy-terminators. Sequencing-PCR products were then purified to remove primers and sequenced with the LICOR NEN model 4300 slab-gel DNA sequencer. The SNP analyses developed in this research were used in two biochemistry laboratory classes. Four novel SNPs identified were: C4198A, found in the NADH dehydrogenase subunit 1 gene; T4586C, T4644G, T4688C 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 HV2 region, however, sequencing was successful for only one of our test subjects. Two previously known SNPs were also found: G3010A and C3116T which are both contained in the 16S rRNA coding region.
Recommended Citation
Punt, Cassidy and Elizabeth Smalley. "Identification of SNPs in the Coding Region of Human mtDNA." Undergraduate Research Symposium, Mankato, MN, April 21, 2008.
https://cornerstone.lib.mnsu.edu/urs/2008/poster-session-B/30
