Sample information |
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| Picture |
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|---|---|
| Location | |
| Collection date | 08/30/2020 |
| Captive / Cultivated? | Wild-caught |
| Group | Bordenstein Lab |
| Observations | A colony of black ants appeared to be living within the tree bark. One was collected at 11:50 am by placing a collection vial over top and allowing it to climb up.. The temperature was 78F, slightly cloudy, and 85% humidity. It is likely a carpenter ant. |
| Putative identification | Arthropoda Hexapoda Insecta Hymenoptera Formicidae Camponotus Camponotus pennsylvanicus |
Methods |
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| Extraction kit | DNeasy (Qiagen) |
| DNA extraction location | Abdomen |
| Single or Duplex PCR | |
| Gel electrophoresis system | MiniOne |
| Buffer | TBE |
| DNA stain | GelGreen |
| Gel images |
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| Protocol notes | This sample was labeled as carpenter ant in the gel. The ant was stored in 70% ethanol for about 3 weeks prior to DNA extraction. DNA Extraction: The ant was incubated in lysis buffer at 56C for 2 hours. Because there was cell debris, I did a 30 sec spin and transferred the supernatant to a fresh tube of 200ul ethanol. This was placed in the freezer overnight and DNA was purified the following day. Eluted DNA was immediately incubated at 65C for one hour prior to PCR. PCR: MiniOne Taq polymerase was used. Gel electrophoresis: The arthropod gel looks whispy; however, that went away as the gel ran longer. This could have been caused by not adding loading dye to samples. The results were very clear, though. I re-colored the gel images in PowerPoint. The MiniOne ladder contains 5 bands: 100, 300, 500, 1000, and 2000 bp. |
Results |
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| Wolbachia presence | Yes |
| Confidence level | High |
| Explanation of confidence level | The DNA extraction was successful because the arthropod COI amplified. Both (+) and (-) controls worked. The Wolbachia 16S rRNA band was present. Sanger sequencing further classified this ant as the black carpenter ant, Camponotus pennsylvanicus, with 100% identity. The Wolbachia strain is Supergroup A. |
| Wolbachia 16S sequence | Download FASTA
Download AB1
TGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCATCCTTAGTTACCATCAGGTAATGCTGGGGACTTTAAGGAAACTGCCAGTGATAAACTGGAGGAAGGTGGGGATGATGTCAAGTCATCATGGCCCTTATGGAGTGGGCTACACACGTGCTACAATGGTGGCTATAATGGGCTGCAAAGTCGCGAGGCTAAGCTAATCCCTTAAAAGCCATCTCAGTTCGGATTGTACTCTGCAACTCGAGTGCATGAAGTTGGAATCGCTAGTAATCGTGGATCAGCACGCCACGGTGAATACGTTCTCGGGTCTTGTACACACTGCCCGTCACGCCATGGGAATTG
BLAST at The Wolbachia Project BLAST at NCBI
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| Arthropod COI sequence | Download FASTA
Download AB1
AATTGGCTCCTCTATAAGAATAATCATTCGACTAGAGTTAGGATCTCCTGATTCACTAATTCTTAATGATCAAACTTTCAATACCATCGTTACAAGTCATGCTTTTATTATAATTTTTTTTATAGTTATACCTTTTATAATTGGGGGATTTGGTAATTTTTTAATTCCACTTATACTAGGATCTCCTGATATAGCTTACCCTCGTTTAAATAACATAAGATTTTGATTACTTCCCCCATCGATCTCCTTATTAATCCTAAGAAATTTTATTAATGAAGGATCTGGAACTGGTTGAACTATCTACCCCCCTCTATCATCAAATACCTTCCATAGTGGCCCCTCTATTGACCTGACTATCTTTTCTCTCCATATTGCTGGTATATCCTCAATTATAGGAGCAATCAATTTTATTTCAACAATTATAAATATACATAATTCCAATATTTCCCTAGATAAAATTCCCTTATTAGTATGATCTATTCTTATTACAGCTATTCTCCTTCTTCTGTCCCTACCTGTTCTAGCAGGCGCTATTACAATACTACTAACAGACCGAAATCTTAATACTTCATTTTTCGATCCCTCGGG
BLAST at The Wolbachia Project BLAST at NCBI
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| Summary | The Camponotus pennsylvanicus was found to be postive for Wolbachia. |
Water Boatman