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Determining Optimal Swab Type and Elution Buffer to Obtain Whole Cells for Future Deconvolution of Complex Cell Mixtures

Jollie, Melissa Lynn
http://rave.ohiolink.edu/etdc/view?acc_num=bgsu161667366449865

Abstract Details

2021, Master of Science (MS), Bowling Green State University, Forensic Science.
An increase of DNA testing sensitivity has led to an elevated variety of sample testing, including complex DNA mixtures, making the interpretation of DNA profiling results more complex. Currently there is no proscribed method used in laboratories to separate complex DNA mixtures by their contributors, therefore a method is needed that could reduce complex mixtures into their component parts. In this study methods of obtaining whole, intact cells from desiccated forensic samples for later cytometric sorting and downstream DNA analysis were examined. This study observed the effects of three different elution buffers (AutoMACS®, Phosphate Buffered Saline, and water) on recovery of whole, intact cells from standard cotton swabs, and then the effect of three different swab types (cotton, flocked, and dissolvable), of different composition, on recovery of whole, intact cells. This was accomplished by washing the swab containing the sample with an elution buffer that maintains the integrity of the cell membrane, resulting in a solution of intact cells. The results of this study demonstrated that the combination of AutoMACS® buffer and flocked swabs provided the highest yield of intact cells post elution. In the future, a solution of whole cells could then be grouped into categories by their cell surface proteins and sorted through cytometric sorting techniques. Once the mixtures have been separated into their component parts by the cell’s surface proteins, then DNA analysis could proceed as normal, potentially resulting in single-source samples.
Crystal Oechsle, Ph.D. (Advisor)
Travis Worst, Ph.D. (Committee Member)
Lewis Maddox, Ph.D. (Committee Member)
36 p.
Biology
Forensic Biology; Whole-cell Elution; Complex Mixture Deconvolution; Cell Sorting

Recommended Citations

Citations

  • Jollie, M. L. (2021). Determining Optimal Swab Type and Elution Buffer to Obtain Whole Cells for Future Deconvolution of Complex Cell Mixtures [Master's thesis, Bowling Green State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu161667366449865

    APA Style (7th edition)

  • Jollie, Melissa. Determining Optimal Swab Type and Elution Buffer to Obtain Whole Cells for Future Deconvolution of Complex Cell Mixtures. 2021. Bowling Green State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=bgsu161667366449865.

    MLA Style (8th edition)

  • Jollie, Melissa. "Determining Optimal Swab Type and Elution Buffer to Obtain Whole Cells for Future Deconvolution of Complex Cell Mixtures." Master's thesis, Bowling Green State University, 2021. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu161667366449865

    Chicago Manual of Style (17th edition)

bgsu161667366449865
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© 2021, all rights reserved.
This open access ETD is published by Bowling Green State University and OhioLINK.