Development of a PCR assay to genotype aromatase knockout mice

CHED 815

Cassandre Noel, cnoel@cse.edu1, Jennifer L. Bogener2, Malaika R. Gallimore2, and Dennis B. Lubahn2. (1) Department of Biochemistry, College of Saint Elizabeth, 2 Convent Rd, Morristown, NJ 07960, (2) Department of Biochemistry, University of Missouri-Columbia, 110A Animal Science Center, Columbia, MO 65211
The aromatase knockout mouse lacks the gene sequence coding for functional aromatase, a protein that converts androgens into estrogens. Previously, to genotype aromatase mice, two PCRs were run: Neomycin and Aromatase. When we attempted to combine both PCRs, the expected band patterns were not attained. To resolve possible factors for the lack of results, we searched the aromatase deleted DNA sequence for possible new primers. Three new pairs of aromatase primers and a neomycin primer pair were chosen. Five PCRs of 15 samples were run using the new and old primers. Following analysis, one new aromatase pair, 2285/2652, was chosen as the best to further experiment. A PCR assay was developed by combining the new and old neomycin primers with the chosen primer, 2285/2652, and two PCRs were run. Since the expected results were attained, 2285/2652 is compatible to run simultaneously with both the new and old neomycin.
 

Undergraduate Research Poster Session: Biochemistry
2:00 PM-4:00 PM, Monday, April 7, 2008 Morial Convention Center -- Hall A, Poster

Division of Chemical Education

The 235th ACS National Meeting, New Orleans, LA, April 6-10, 2008