Date of Award

12-1-2004

Degree Type

Restricted Access Professional Report

Degree Name

Master of Science

Field of Study

Cell Biology and Genetics

Department

Graduate School of Biomedical Sciences

First Advisor

John Planz

Second Advisor

Joseph Warren

Third Advisor

Arthur Eisenberg

Abstract

Amersham Bioscience produces the only available WGA kit, GenomiPhi DNA Amplification kit. It is a multiple displacement amplification reaction. The kit contains all reagents needed to perform WGA. Optimal amount of DNA input into the reaction is 1 ng or greater to yield 3 μg product. This amount is the optimal amount currently needed by PCR systems; however STR amplification requires the 1 ng in 10 μL of sample extract. GenomiPhi requires 1 μL of sample extract, ten times the concentrations of STR systems. The goal of this project was to evaluate the possible uses of the GenomiPhi DNA Amplification kit for use in the forensic community. The ability to amplify the entire genome without bias would benefit low copy number samples where there is little DNA to start with. One objective of the study was to determine the lower limit of input DNA into the GenomiPhi reaction. Input DNA varied from 10 ng to 7 pg. Secondly, input DNA was degraded to determine how the GenomiPhi kit will be affected by the input of less than pristine DNA. All samples were quantitated by PicoGreen assay, STR amplified with Profiler Plus and analyzed on ABI’s 310 Genetic Analyzer. Samples were analyzed before and after WGA to determine under which circumstance better results were seen.

Comments

W 4.8 H196E 2004

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