Kris Rusinko

Document Type

Honors Project

First Advisor

Dr. Stephen Baron

Degree Award Date

Winter 2000

Keywords

PCR Reaction, Chicken Samples, Salmonella

Disciplines

Bacteria | Laboratory and Basic Science Research | Medicine and Health Sciences

Abstract

The ultimate goal of this lab was to qualitatively test chicken samples for the presence of salmonella bacteria by PCR methods. However, this lab is only the preliminary workings in determining the specificity of the FimAF and FimAR primers designed at the beginning of the experiment to cut and amplify the FimA gene specific to salmonella bacteria. Known strains of salmonella bacteria DNA were isolated and amplified through PCR techniques. These resultant DNA mixes were then run through electrophoresis gels versus DNA ladders to check for the presence of the FimA gene by searching for its 453 base pair size band on the gel. This was run versus several controls to test the specificity of the FimA gene to the salmonella and in an attempt to be certain that it was not present in other DNA samples. Very strong bands were found in the salmonella DNA samples, and only very weak bands in any of the other samples. This band was then cut out of one of the gels and a Silver Staining method for DNA sequencing was performed to be certain that it was the FimA gene that was amplified. Upon inspection of the resulting sequence, a 120 base pair sequence that matched that of the FimA gene was located. This proves that the FimAF and FimAR primers do amplify the FimA gene and work for salmonella testing

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