Date of Award
Restricted Access Professional Report
Master of Science
Field of Study
Cell Biology and Genetics
Graduate School of Biomedical Sciences
Species identification is not only important in determining the origins of remains found in human forensic cases, but is also important in the growing field of nonhuman forensics. Animal forensics is an emerging discipline in the non-human forensics field. Animal forensics focuses on domestic animals, or animals that live in close contact with humans. These animals include dogs, cats, cattle, pigs, horses, sheep as well as others. Objective 1: Sensitivity - Forensic samples are often low copy number so there may be little viable or non-degraded DNA to work with. The PCR reaction needs to be optimized to determine how little DNA you can start with and still obtain accurate results. The given protocol from DNA Solutions, Inc. (Appendix A) [ 17] suggests 20ng of input DNA. In practice, this level is rarely found in forensic cases, so this assay will begin at 5ng of 9 input DNA. The dilution series will be 5ng, 2ng, 1 ng, 500pg, 250pg, 125pg, 62.5pg, 31.25pg and 15.63pg. Objective 2:PCR cycle number- As a part of sensitivity, PCR cycle number is an important part of optimizing the reaction. Since agarose gels are being run to visualize results, increased cycle number can result in increased specificity. Cycle numbers being evaluated are 26 cycles [ 17], 28 cycles, 30 cycles, 32 cycles and 34 cycles. Objective 3: Species Specificity - It must be determined if these universal primers do, in fact, bind to all species that possess the genes of interest. Samples from across the different animal classes will be evaluated to determine the specificity of the primers. The primers have already been tested on members of the deer family, with success, so a male deer sample has been sent to be used as a positive control as it shows both genes. Species of interest are mammals, birds and fish as there are many of each that are endangered and identifying the sex of the animal is an important step to identifying the individual animal.
Price, G. N.
"Evaluation of Unlabeled Primers for Sex Determination of Animal and Wildlife Samples" Fort Worth, Tx: University of North Texas Health Science Center;