ABSTRACT
INTRODUCTION
The majority of eukaryotic organisms have a genetic composition that is unique in comparison to other living organisms (Goldstein 2003). Less unique, with only limited variation, is the shared genetic composition of the species within which each organism belongs to. Efficient identification of organisms based on their genetic composition is available due to current techniques for tissue sampling and statistical analysis (Avise, Nelson & Prodohl 1996). Application of molecular genetic assays was utilized to identify a sample of tissue with an unknown identity. Initially, deoxyribonucleic acid (DNA) was extracted and assayed in an electrophoretic gel for necessary content (Campbell, Harriss, Elphinstone, & Baverstock 1995).…show more content… Each tube received 5.5 μL ddH2O. Added to a single tube was 0.5 μL forward primer (ND4) while 0.5 μL reverse primer (Leu) was added to the remaining tube. Next, 2.0 μL clean PCR product and 2.0 μL Big Dye Terminator Cycle Sequencing Buffer was added to each tube respectively. Thermal cycling was then completed by the lab instructor using GeneAmp® PCR system 9700 paired with protocol of Texas State University Genetics: 2450 Laboratory Manual (Forstner et al. 2013). The CSR consists of 40 cycles, each of which consists of 20 seconds at 96°C for denaturation, 20 seconds at 50°C for primer annealing, and four minutes at 60°C for primer extension. CSR products were then stored at 4°C prior to CSR clean…show more content… 2013). The DNA sample was then precipitated and washed free of contaminants. Following extraction the sample failed electrophoretic gel assay for success prior to target DNA amplification. The extraction failure may have been do to errors in any of the extraction technique applications. The absence of any fluorescence on the electrophoretic gel assay (Figure 1) suggests the DNA sample was not successfully eluted from the silica gel membrane or the sample was deposited too deep in the agarose gel prior to the application of electrical current causing it to fall below ultraviolet light
