Sample information |
|
| Picture |
|
|---|---|
| Location | |
| Collection date | 02/19/2025 |
| Captive / Cultivated? | Captive / Cultivated |
| Group | Berkshire Community College |
| Observations | |
| Putative identification | Arthropoda Insecta Blattodea Blaberidae Blaptica Blaptica dubia |
Methods |
|
| Extraction kit | Monarch DNA extraction (NEB) |
| DNA extraction location | Rear half |
| Single or Duplex PCR | Duplex Reaction |
| Gel electrophoresis system | Edvotek Gel Electrophoresis |
| Buffer | TAE |
| DNA stain | SYBR Safe |
| Gel images |
|
| Protocol notes | For DNA extraction we did cell lysis, DNA binding and purification, and DNA Elution. Taq polymerase used: New England Biolabs OneTaq Hot Start Quick-Load 2x MM w/ Standard Buffer. First PCR set up on 3/5/25 and was a single PCR with only Arthropod primers and the annealing temperature was 49 C. The second PCR Reaction set up date 3/26/25 and was a duplex PCR with both Arthropod and Wolbachia primers and the annealing temperature was 49 C. DNA Ladder used: New England Biolabs 1 kb Plus DNA Ladder for Safe Stains. |
Results |
|
| Wolbachia presence | Unknown |
| Confidence level | Medium |
| Explanation of confidence level | Arthropod DNA didn’t show up but the controls worked. Next time would add more DNA for the Arthropods. |
| Wolbachia 16S sequence | |
| Arthropod COI sequence |
|
| Summary | |
Common Eastern Bumble Bee (Bombus impatiens)
American Bird
Spotted crane fly
Wolbachia data
Meadow Katydid