Bee (Bombus sp. StKate18-WP4)

November 5, 2022

Sample information
Picture
Photos by: Elizabeth K.
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
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
Protocol notes

Results
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
Summary The Hymenoptera was found to be negative for Wolbachia.
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