Title

Who Done It?: DNA Identification of Species-Specific Samples Using Real Time Polymerase Chain Reaction

Faculty Sponsor

Dr. Beth Scaglione-Sewell

College

Arts and Sciences

Department/Program

Biology

ORCID Identifier(s)

0000-0002-8168-7507, 0000-0002-9034-3458

Document Type

Poster Presentation

Symposium Date

Spring 4-23-2016

Abstract

The cytochrome oxidase subunit I (COX1) gene has been used for many years to identify invertebrates and vertebrates. For example, COX1 is utilized to identify bird (avian) species and forensically important blowflies (Calliphoridae) by their DNA. The traditional method is to use polymerase chain reaction (PCR) on a sample and to subsequently sequence the COX1 gene. Because it is not cost-efficient for small research laboratories to sequence numerous samples at one time, the goal of these experiments is to develop an alternative method to DNA sequencing for identification of species from DNA samples using real-time polymerase chain reaction (RT-PCR). Through a genomic DNA preparation and gene amplification using PCR, the COX1 DNA region was taken from avian fecal samples and from Calliphoridae samples. Nine Calliphoridae COX1 DNA were taken from Calliphoridae maggot samples and successfully verified via sequencing to be Lucilia silvarum, Calliphora vicina, and seven Lucilia Illustris. Avian COX1 DNA from fecal matter has been detected and amplified. Future work includes using RT-PCR with DNA samples and developing species-specific primers for the COX1 gene. Calliphoridae COX1 DNA sequences are extremely homologous with only slight variations, thus, requiring the use of species-specific fluorescent probes. Avian COX1 DNA sequences differ greatly between species allowing for the use of SYBR® green with species-specific primers.

Biographical Information about Author(s)

Max Carpenter is a senior biology major. He will be attending Marquette University School of Dentistry next year. He is also the president of Valparaiso University Pre-Medical Arts Club. Ryan Banashak is a senior biology, chemistry double major. He is in the process of applying to medical school. He is currently a medical scribe hoping to expand his knowledge for medical school. Rahul Rayan is a junior biology, chemistry double major with a psychology minor. He is also in the process of applying to medical schools.