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Ursidae: The Undergraduate Research Journal at the University of Northern Colorado

Faculty Sponsor

Gregory DeKrey

Faculty Sponsor Email

gregory.dekrey@unco.edu

Abstract

The goal of this study was to determine if flow cytometry could be used to identify transgene carrier mice [B6.Cg-Tg(Prdm1-EYFP)1Mnz/J] by their expression of the enhanced yellow fluorescent protein (eYFP) in peripheral blood lymphocytes. In these mice, eYFP expression is under the control of the Prdm1 gene promoter. Littermates were identified as being either wild type or transgene carriers using a polymerase chain reaction (PCR) analysis of tail tissue. Peripheral blood leukocytes were isolated from each of the mice, and both the percent and mean fluorescence intensity (MFI) were determined for eYFP expression by lymphocytes. We found that blood lymphocytes from eYFP transgene carrier mice contained an average of 1.54% eYFP+ cells. In contrast, wild type animals contained significantly fewer (P < 0.05) with an average of 0.18%. The percent eYFP- and eYFP+ data ranges did not overlap. The average MFI values for these groups were also significantly different, but their data ranges overlapped. The PCR process performed in our laboratory required approximately 20 hours over three days to complete with an estimated per animal cost of $16.00. In contrast, the flow cytometry procedure required approximately six hours within one day to complete with an estimated per animal cost of $7.85. We conclude that flow cytometry is an adequate and less costly method for identifying eYFP transgene carrier mice.

UNCO Undergraduate Verification

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