Sample information |
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| Picture |
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|---|---|
| Location | |
| Collection date | 08/23/2021 |
| Captive / Cultivated? | Wild-caught |
| Group | The School for Science and Math at Vanderbilt |
| Observations | This white-fringed weevil was caught on the underside of a wooden mailbox. It was in the shade under a tree, in a neighborhood. |
| Putative identification | Arthropoda Hexapoda Insecta Coleoptera Curculionidae |
Methods |
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| Extraction kit | DNeasy (Qiagen) |
| DNA extraction location | Abdomen |
| Single or Duplex PCR | Single Reaction |
| Gel electrophoresis system | MiniOne |
| Buffer | TBE |
| DNA stain | GelGreen |
| Gel images |
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| Protocol notes | *PCR machine failed during DNA amplification for 7 test tubes; the Bordenstein lab reran our samples. *We identified this arthropod as a white-fringed weevil based on morphology and what BLASTN results suggested, but chromatogram results for CO1 were not completely clear, therefore our identification is not conclusive. |
Results |
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| Wolbachia presence | Yes |
| Confidence level | High |
| Explanation of confidence level |
High confidence level is due to appropriate (expected) test results from controls in both CO1 and WSPEC experiments. Positive controls were positive, negative control and water were Wolbachia negative, water was negative for arthropod DNA. There did not appear to be contamination between samples. |
| Wolbachia 16S sequence |
GTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCATCCTTAGTTGCCATCAGGTAATGCTGAGCACTTTAAGGAAACTGCCAGTGATAAGCTGGAGGAAGGTGGGGATGATGTCAAGTCATCATGGCCTTTATGGAGTGGGCTACACACGTGCTACAATGGTGTCTACAATGGGCTGCAAAGTCGCGAGGCTAAGCTAATCCCTAAAAAACATCTCAGTTCGGATTGTACTCTGCAACTCGAGTACATGAAGTTGGAATCGCTAGTAATCGTGGATCAGCATGCCACGGTGAATACGTTCT
BLAST at The Wolbachia Project BLAST at NCBI
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| Arthropod COI sequence |
TTCGTTCAGAATTAGGAAACCCNGGATCTCTGTTGGAGATGACCAAATTTATAACGTTATTGTAACTGCTCATGCTTTTA TTATAATTTTCTTTATAGTAATACCTATCATAATTGGGGGATTTGGAAACTGACTAGTTCCATTAATACTAGGAGCTCCA GACATAGCCTTCCCTCGTCTTAATAATATAAGATTTTGACTACTTCCACCTTCTTTAACTTTACTTTTAATAAGAAGAAT TGTTGATAAAGGTGCAGGAACAGGATGAACAGTNTATCCCCCTCTCTCTGCCAATGTAGCTCATGAAGGATCCTCTGTTG ATTTAGCAATTTTTANACTACATATAGCAGGTGTATCATCAATTTTAGGAGCAGTAAATTTTATTTCTACAGTTTTAAAT ATACGACCTTCAGGAATATCTCCTGATCGAATACCTTTATTTGTATGAGCAGTAGAAATTACTGCTATNTTTATTACTTT TATCTTTACCAGTTTTAGCAGGAGCAATNACTATACTTTNTAACAGATCGAAATATTAATACCTCATTTTTTGACNCCTG CAGGAGGAGGAGATCCTATTTTATANCAACATTTATTTTGATTTTTTGG
BLAST at The Wolbachia Project BLAST at NCBI
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| Summary | The Curculionidae was found to be postive for Wolbachia. |
Water Boatman