Who Done It?: DNA Identification of Species-Specific Samples Using Real Time Polymerase Chain Reaction
Dr. Beth Scaglione-Sewell
Arts and Sciences
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. Calliphoridae COX1 DNA was taken from two Calliphoridae maggot samples and successfully verified via sequencing to be Lucilia silvarum and Calliphora vicina. Further testing must be done to detect avian COX1 DNA from fecal matter. 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.
Carpenter, Max and Hargrave, Lauren, "Who Done It?: DNA Identification of Species-Specific Samples Using Real Time Polymerase Chain Reaction" (2015). Symposium on Undergraduate Research and Creative Expression (SOURCE). 484.