Sample information |
|
| Picture |
|
|---|---|
| Location | |
| Collection date | 02/18/2025 |
| Captive / Cultivated? | Captive / Cultivated |
| Group | Berkshire Community College |
| Observations | The crickets lived in a box in Petco altogether. |
| Putative identification | Arthropoda Hexapoda Insecta Orthoptera Gryllidae Acheta Acheta domesticus |
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 could have incubated for longer than we did. Incubation time was 1 minute and we didn’t set an exact timer. Other than that, the protocol was followed. The Taq polymerase used were the New England Biolabs One Taq Hot Start. For PCR with the arthropod primers the annealing temp was 49C. Updated: For the duplex PCR the annealing temp was 49C. The Taq polymerase used-NEB OneTaq Hot Start Quick-Load 2x MM w/stand. Date we set up second PCR reaction is 3/26/25. Updated 4/2/25: Taq polymerase used: New England Biolabs OneTaq Hot Start Quick-Load 2x MM w/ Standard Buffer DNA Ladder Used: New England Biolabs 1 kb Plus DNA Ladder for Safe Stains |
Results |
|
| Wolbachia presence | No |
| Confidence level | High |
| Explanation of confidence level | Great controls and results that are very clear. Very high confidence that our arthropods are Wolbachia negative. |
| Wolbachia 16S sequence | |
| Arthropod COI sequence |
|
| Summary | The Acheta domesticus was found to be negative for Wolbachia. |
Baetis_C2.2
Formica_C11
Philaenus sp._C8
Ichneumonidae_A12
Formica_A11