Sample information

Caught on a concrete path, with grass on each side of it close to a building. The concrete is a light color. Located in a woody, suburban area, with many types of different vegetation. 

Caught late summer/early starts of fall

Temperature: High of 82℉; Low of 62℉

Terrestrial Habitat

Color: The arthropod is a rusty reddish brown color, with some areas being lighter than the others. Additionally, compared to it when it was first caught, it appears that it lost some of its color after being in the alcohol. 

Wings: No wings

Legs: This arthropod has 6 legs, with 3 on each side. The forelegs are connected to the thorax and look to be a little shorter than the others, however, it was hard to tell since the arthropod curled up after being in the freezer and alcohol. The legs look to be segmenting, with certain areas having joints where the legs bend. The hind legs are very similar to the fore legs, with the only difference being that they look to be proportional to the body length and they are the longest legs out of the 3 pairs. 

Antennae: Has two antennae, which are elbow shaped. In other words, they are divided into what looks to be 2 segments, with one segment seeming longer than the other. 

Body: There are 3 apparent body regions: the head, thorax, and abdomen. Each of the segments seems to be rounded. The head has a pair of mandibles, antennae, eyes, and a really small mouth that is hard to see. The thorax and abdomen seem to be connected by a small waist type situation containing two different sections. The body is covered by an exoskeleton. Additionally, the abdomen looks to have something maybe a little pointy or sharp on it, however, it can be part of the exoskeleton.

Methods

DNA Extraction: 

For this process, all of the arthropods were used. During this, this sample did have some type of bubbles before we incubated the sample, though it wasn’t as much as other samples. Additionally, this sample solution was a tanish color. 

PCR:

For the PCR process, we did a CO1 one and a Wolbachia one. In the overall PCR process, there was a mistake where the DNA of 2 of the 6 samples was not added. However,  this particular sample wasn’t one of those, leading to no known mistakes for A-6. 

Gel electrophoresis, Arthropod CO1:

We used two different gel electrophoresis machines. 

For machine 1:

Lane 1 – DNA Ladder 

Lane 2 – Arthropod A-1 

Lane 3 – Arthropod A-2 

Lane 4 – Arthropod A-3

Lane 5 – Arthropod A-4

Lane 6 – Wolbachia + control

Lane 7 – Wolbachia – control

Lane 8 – Water 

Lane 9 – Blank 

For machine 2: 

Lane 1 – DNA Ladder 

Lane 2 – Arthropod A-5 

Lane 3 – Arthropod A-6 

Lane 4 – Wolbachia + control

Lane 5 – Wolbachia – control

Lane 6 – Water 

Lane 7 – Blank 

Lane 8 – Blank

Lane 9 – Blank

All of the controls worked, with the Wolbachia (+) control and Wolbachia (-) control having the CO1 band and the water control having no band, meaning no contamination was made. Meaning that any mistakes made, in other words no bands showing up for arthropod sample, was because of an error in either the PCR process or loading the gel electrophoresis machine. This sample did have a band show up that was readable and in the correct location, being at 708bp. However, we continue with the results from the gel since the band was at the correct location.

Gel electrophoresis, Wolbachia:

We used two different gel electrophoresis machines. 

For machine 1:

Lane 1 – DNA Ladder 

Lane 2 – Arthropod A-1 

Lane 3 – Arthropod A-2 

Lane 4 – Arthropod A-3

Lane 5 – Arthropod A-4

Lane 6 – Wolbachia + control

Lane 7 – Wolbachia – control

Lane 8 – Water 

Lane 9 – Blank 

For machine 2: 

Lane 1 – DNA Ladder 

Lane 2 – Arthropod A-5 

Lane 3 – Arthropod A-6 

Lane 4 – Wolbachia + control

Lane 5 – Wolbachia – control

Lane 6 – Water 

Lane 7 – Blank 

Lane 8 – Rerun Arthropod A-1

Lane 9 – Rerun Arthropod A-2

The reruns at the end of the machine 2 gel electrophoresis were because no CO1 band showed up on that gel for those samples due to the DNA being forgotten in the PCR process. All of the controls worked, with the Wolbachia (+) control having a Wolbachia band at 438bp  and the Wolbachia (-) control and the water control having no band, meaning no contamination was made. Meaning that any mistakes made, in other words no bands showing up for arthropod sample, was because of an error in either the PCR process or loading the gel electrophoresis machine. This sample did have a Wolbachia band that showed up, meaning that this arthropod was positive for Wolbachia.

Results

My confidence level is what it is because everything seemed to go according to plan with the process. With this, all of the controls worked properly and there were no known mistakes made in the process of this sample. So, I’m pretty confident with this arthropod’s results.