Developing a QPCR Genotyping Method to Screen for Apolipoprotein E Variants

Presenter Information

Jessica Jurovich, Minnesota State University, Mankato

Location

CSU Ballroom

Start Date

9-4-2012 10:00 AM

End Date

9-4-2012 11:30 AM

Student's Major

Chemistry and Geology

Student's College

Science, Engineering and Technology

Mentor's Name

James Rife

Mentor's Department

Chemistry and Geology

Mentor's College

Science, Engineering and Technology

Description

A less time-intensive, real-time polymerase chain reaction (qPCR) genotyping method to screen for apolipoprotein E variants was developed. Certain apolipoprotein E variants have been associated with increased risk of Alzheimer’s disease and hypolipoproteinemia. Using the DNA sequences for each variant found on the National Center for Biological Information website, the sites of variation were determined, and primers and probes that can discriminate between the E2, E3 and E4 alleles of apolipoprotein E were developed. The probes were fluorescently labeled and designed to selectively bind to specific DNA variants. Buccal samples from several volunteers were obtained. DNA was isolated from these samples using a QIAamp DNA mini Kit from QIAGEN. A Step One Plus real-time PCR system from Applied Biosystems was then used to determine the genotypes by monitoring which probes were used more rapidly for each sample. This procedure will be used as an experiment in future CHEM 360 labs.