Sample information

Red Fire Ants are pretty small so this did concern me if the DNA would be effectively amplified to be shown in the end results. But it’s size and body structure made DNA extraction fairly easy.

Methods

Lane 1: DNA ladder, Lane 2: CO1 gene from insect genomic DNA (Fire Ant 1) Lane 3: CO1 gene from insect genomic DNA (Fire Ant 2), Lane 4: Wolbachia DNA (Fire Ant 1), Lane 5: Wolbachia DNA (Fire Ant 2) Experimental Set Up

• Sample Collection, two Red Fire Ants were placed into tubes
• Cell Lysis, cell lysis buffer was added to each tube, mash for 1 minute, add another 100 µL, and mash again.
• Heat Treatment, tubes were sealed with parafilm and heated for 5 minutes.
• DNA Precipitation Setup, added cold isopropanol to each tube for DNA precipitation.
• Cooling and Centrifugation, samples were cooled to room temperature, vortexed, then centrifuged.
• Supernatant Transfer, transferred 150 µL of supernatant to new tubes with isopropanol, then centrifuge for 5 minutes.
• Washing, supernatant was discarded, pellets air dried (DNA), then ethanol was added and centrifuged
• Arthropod PCR of Insect 15, Arthropod PCR of Insect 16, Wolbachia, PCR of Insect 1, Wolbachia PCR of Insect 2 were placed into gel loading tray and into the electrophoresis machine

Results

My results indicated that Wolbachia was present because the results from gel electrophoresis showed bands with the corresponding size. Rows three and four of the Wolbachia DNA had clear bands indicating it was found. When comparing my data to my control group they looked similar so I am confident it was successful that Wolbachia was found in both Red Fire Ants.