Extraction and quantification of human deoxyribonucleic acid, and the amplification of human short tandem repeats and a sex identification marker from fly larvae found on decomposing tissue

Keywords

DNA -- Analysis, Forensic sciences, Polymerase chain reaction

Abstract

The purpose ofthis experiment was to determine if human deoxyribonucleic acid (DNA) could be extracted, quantified, and amplified from fly larvae that had fed on decomposing human tissue. Fly eggs were collected and placed on donated human skin, muscle, or liver tissue in rearing chambers. After two to twelve days, the larvae were collected in 70% ethanol, 2-propanol, acetone, or by freezing. One to sixteen larvae were dissected and the DNA extracted using digest buffer and either phenol: chloroform: isoamyl alcohol or by using silica spin columns. The human DNA was quantified using a biotinylated primate specific probe. The DNA extract was then amplified using the polymerase chain reaction (PCR) and primers for 13 specific Short Tandem Repeats (STRs) and the sex identification marker amelogenin. Finally, the PCR product was separated and characterized using capillary electrophoresis. Out ofthe 30 DNA extracts, one sample had approximately 4.5-9.0ng of human DNA. The remaining 29 extracts were either void of human DNA or the amount ofhuman DNA fell below the test detection limits. Three ofthe thirty extracts produced amplified products for the sex identification marker and seven to eight STRs. The STR profiles produced by these three extracts matched the STR profiles ofthe corresponding positive controls. The experiment also provided evidence, but not conclusive proof, that the human DNA originated from inside the larvae.

Notes

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Graduation Date

2001

Advisor

Tilstone, William J.

Degree

Master of Science (M.S.)

College

College of Arts and Sciences

Department

Chemistry

Format

PDF

Pages

113 p.

Language

English

Length of Campus-only Access

None

Access Status

Masters Thesis (Open Access)

Identifier

DP0028713

Subjects

Arts and Sciences -- Dissertations, Academic; Dissertations, Academic -- Arts and Sciences

Accessibility Status

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