Analysis of mitochondrial DNA from an ancient Miami Indian : an honors thesis (HONRS 499)

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Cummings, Christopher M.
Vann, Carolyn N.
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Thesis (B.?.)
Honors College
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The purpose of this research study was to analyze the mitochondrial DNA of an ancient Miami Indian. The results from this study may be used by other researchers to make comparisons to other individuals of the Miami Indian population, ancient and modem. The Miami people lost federal recognition in the mid-1800's. If a close genetic relationship between a modem and known ancient population can be proven, a strong case may be available for regaining the recognition of these people by the federal government.For this analysis, a tooth was provided from a burial site that was excavated in Henry County, IN. The tooth has been stored in the Anthropology department's museum and has been donated by Don Cochran for use in this project. The Ancient DNA was extracted from the dentin of the tooth that was drilled by Dr. Neal Lambert DDS. The DNA was cut with a blunt-ended restriction enzyme, Hae III. Double-stranded DNA adaptors were ligated to the blunt ends. A single primer was used to amplify the resulting fragments using PCR. Using this library, the DNA was readily reamplified using a small amount of the PCR product. The individual's haplotype was determined by amplifying specific regions of its mtDNA. An agarose gel was created to visualize any existing amplification. Upon successful amplification, sequencing of the individual's D-loop may be possible to categorize the individual into a haplogroup.Amplification was evident based on observations from the agarose gel, but contamination from DNA other than this individual's was present. Several amplifications and agarose gels were created to seek evidence to determine this individual's haplotype, but were ultimately unsuccessful. Sequencing may be an option after future procedures are undertaken to create a successful amplification and agarose gel. Contamination would need to be eliminated in order to sequence the DNA, most likely achieved by a more cautious application of methods.