Sample information |
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| Picture |
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|---|---|
| Location | |
| Collection date | 09/14/2022 |
| Captive / Cultivated? | Wild-caught |
| Group | St. Albans School |
| Observations | I found this bee pollinating a flower in the NCS middle school amphitheater. There were many other bees in the area. It was collected at 10:30am by placing a ziplock bag around the flower the bee was pollinating and slowly capturing the insect by removing it from the flower and into the bag. The bee was buzzing, active, loud, and visibly fuzzy. It was killed in rubbing alcohol and stored in a freezer for about 3 hours, until it was later used. |
| Putative identification | Arthropoda Hexapoda Insecta Hymenoptera |
Methods |
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| Extraction kit | DNeasy (Qiagen) |
| DNA extraction location | Abdomen |
| Single or Duplex PCR | Single Reaction |
| Gel electrophoresis system | MiniPCR |
| Buffer | TBE |
| DNA stain | GelGreen |
| Gel images |
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| Protocol notes | |
Results |
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| Wolbachia presence | No |
| Confidence level | Medium |
| Explanation of confidence level | The DNA extraction was successful as all gels and controls for arthropod DNA were apparent as bands. At least one of both Wolbachia controls were executed correctly and worked. However, one (-) Wolbachia control was contaminated with moth DNA and one (+) DNA control band did not work. |
| Wolbachia 16S sequence | |
| Arthropod COI sequence | Download FASTA
Download AB1
AGGATCATCTATAATATTATTAATTCTAATAGAACTTATACATCCAGGAATATGAATTAATAATGATCGAATTTATAACTCATGATTTACTAGACGTGCATTTTTAATAATTTTTTTTATAGTTATACCATTTATAATTGGAGGATTTGGAAATTATTTAATTCCTTTAATATTAAGATCACCTGATATAGCTTTTCCACGAATAAATAATATTAGATTTTGATTATTACCTCCATCTATTTTTATATTATTATTAAGAACTTTATTTACACCTAATGTAGGAACAGGTTGAACCGAATATCCCCCCTTATCATCTTATTTATTTCATTCNTCACCTTCNNTAGATATTGCAATTTTTTCTTTACATGTAACNNGAATTTCTTCTATTATTGGNTCATTAAATTTTATTGTTACTATTATAGTAATAAAAAATTTTTCNTTAAATTATGATCNNATTACTTTATTTTCATGATCTGNNNGNNTTACAGTATTATTATTAATTTTATCATTACCNGTTNCANGNGGAGCAATTACTANNCTTCTTTNTGATCGANANTNTAATACATCATTNTTTGATCCANTAGGAGGCGGAGATCCAATATNTATATCNAC
BLAST at The Wolbachia Project BLAST at NCBI
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| Summary | The Hymenoptera was found to be negative for Wolbachia. |
Water Boatman