Sample information

Black ant; about 5mm; 6 legs; redish colored legs and antennae; hairs on body; head, thorax, and abdomen are shiny black; fast runner. The common name for the ant is a black field ant.

I found the ant in the “backyard” of a school at a clearing before the entrance to the woods. The ant was collected in late May, the temperature was 20.556º C, it was sunny, and the ant was crawling across a patch of dry dirt among some weeds and grass.

Methods

DNA extraction: I simply mashed the entire bug to collect the abdomen DNA

PCR: I made a mistake during PCR because I pipetted too much “PCR cocktail” into one of the PCR tubes (not sure which) and didn’t have enough cocktail for the Wolbachia control (it is omitted from the gel).

Gel electrophoresis lanes for the image above:

1. DNA ladder 1
2. My ant tested for CO1 arthropod gene
3. My lab partner’s ant tested for CO1 arthropod gene
4. Positive arthropod control
5. Negative arthropod control
6. Blank (was supposed to be positive wolbachia dna control)
7. DNA water control
8. Break
9. My ant tested for wolbachia dna
10. My lab partner’s ant tested for wolbachia dna
11. Positive arthropod control
12. Negative arthropod control
13. Positive wolbachia control
14. Water control
15. DNA ladder 2

Analysis:

The ants had very strong bands, but the controls did not work as expected. Additionally, the wolbachia side of the gel has faint bands at the arthropod bp location.

Sequencing: Unfortunately, my wolbachia sequencing sample dried up and was unable to be sequenced. Only the arthropod sample was sequenced, resulting in an average Q-value of 39. When I “blastn” and “blastx” the sequence, I got my type of arthropod.

Results

I am somewhat confident my ant is wolbachia positive because although the band is extremely bright at the wolbachia base-pair location on the gel (comparing with the DNA ladder), the negative wolbachia control bug had a faint band at the wolbachia base-pair location. So, I’m not completely confident in my results but I am fairly confident. Additionally, my wolbachia sample was unable to be sequenced, so I don’t actually know for sure if my ant had wolbachia. Nevertheless, the highlighted bands give me confidence in my results.