Sample information

Found under rotting wood on damp soil with colony of other isopods of similar appearance. It was caught at around 4:40 pm, and the temperature outside was 5°C.

Methods

1. DNA Extraction – This isopod was quite small and its exoskeleton was fairly soft. We cut its abdomen in half and scored the exoskeleton with the scalpel to break it up without damaging the rest of its abdomen. It got crushed very well and did not contain any floating parts at the end.

Gel Electrophoresis – For CO1 arthropod

The first two wells got damaged so we had to move over:

3: NEB NO551L Ladder

4: Isopod #1

5: Isopod #2

6: Isopod-R #1

7: Isopod-R #2

8:

9:

10: Positive Control

11: Negative Control

12: DNA Postive Control

13: Water / DNA Negative Control

Wolbachia 16S

1. NEB NO551L Ladder

2. Isopod #1

3. Isopod #2

4.

5.

6. Isopod-R #1

7. Isopod-R #2

8.

9.

10. Positive Control

11. Negative Control

12. DNA Positive Control

13. Water / DNA Negative Control

Analysis: Both positive and negative controls did not work. During Gel Electrophoresis, Isopod #1 had a band for Arthropod CO1 and did not have a band for Wolbachia 16S, but these results cannot be trusted. Most likely there was an error somewhere in the DNA Extraction and PCR steps.

Results

Our water (DNA – control) was contaminated with DNA, and our Wolbachia positive control did not indicate presence of Wolbachia. Therefore our confidence level is low and our results cannot be trusted.