Sample information

I found this woodland fuzzy ant underneath a dead wood log that I flipped. Next to this ant were dozens of ant larvae, indicating the existence of a nest nearby. It was during the spring at ten in the morning, and it had just finished raining, thus everything was wet. I located the log next to a river bank and it was roughly 68 degrees F. There were many porthole ants but I took an interest to this one as it seemed like it had wings.

Methods

Gel Image Identification:

571, 572 (lasius americanus), 573, 574, Positive Wolbachia Drosophila, Negative Drosophila, PC DNA, Water, Ladder

The bottom row shared the same samples, but for Wolbachia primers. L to R:

571, 572 (lasius americanus), 573, 574, Ladder, Positive Wolbachia Drosophila, Negative Drosophila Drosophila, PC DNA, Water

Primers to specifically amplify a 438-bp (base pair) fragment of the 16S ribosomal RNA gene (ubiquitous in all Wolbachia) are:
Wolbachia_F (16S_WspecF): 5’−CAT ACC TAT TCG AAG GGA TAG−3’  and
Wolbachia_R (16S_WspecR): 5’−AGC TTC GAG TGA AAC CAA TTC−3’.
When using these primers, the ideal annealing temperature is 55°C.

Primers to specifically amplify a 708-bp fragment of the CO1 cytochrome oxidase gene (ubiquitous in arthropod mitochondria) are:
Arthropod_F (LCO1490): 5’−GGT CAA CAA ATC ATA AAG ATA TTG G−3’  and
Arthropod_R (HCO2198): 5’−TAA ACT TCA GGG TGA CCA AAA AAT CA−3’.
When using these primers, the ideal annealing temperature is 49°C.

Results

For my group’s Wolbachia gel, our positive drosophila control (which was supposed to show up) did not appear, which made us suspect a false negative error for all the other samples as well.