Development of Real-Time Polymerase Chain Reaction (q-PCR) Assays for the Specific Detection and Characterization of Select Bacterial Pathogens

Authors

Dr. Richard Robison, Brigham Young University

Keywords

polymerase chain reaction, q-PCR, bacterial pathogens

College

Life Sciences

Department

Microbiology and Molecular Biology

Abstract

Primer generation software will be used to design primer and probe sequences that will theoretically perform optimally in PCR reactions. All sequences will be subjected to BLAST searches to confirm their specificity for a particular organism. Once primer sequences have been obtained, they will be synthesized by Integrated DNA Technologies, Inc. (IDT). These primers will be evaluated in q-PCR reactions using target DNA from multiple isolates and SyberGreen to detect nucleic acid amplification. Parameters for each reaction will be optimized using these same tools. Once a reaction has been optimized, a specific labeled probe will be synthesized which contains both a fluorescent dye and its specific quencher. This will be used in a Taqman® PCR reaction to detect specific target sequence amplification. The following q-PCR reactions will be developed:

Recommended Citation

Robison, Dr. Richard (2013) "Development of Real-Time Polymerase Chain Reaction (q-PCR) Assays for the Specific Detection and Characterization of Select Bacterial Pathogens," Journal of Undergraduate Research: Vol. 2013: Iss. 1, Article 1818.
Available at: https://scholarsarchive.byu.edu/jur/vol2013/iss1/1818