Sample information |
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Picture |
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Location | |
Collection date | 02/18/2025 |
Captive / Cultivated? | Captive / Cultivated |
Group | Berkshire Community College |
Observations | |
Putative identification | Arthropoda Hexapoda Insecta Diptera Drosophilidae Drosophila Drosophila melanogaster |
Methods |
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Extraction kit | Monarch DNA extraction (NEB) |
DNA extraction location | Whole arthropod |
Single or Duplex PCR | Single Reaction |
Gel electrophoresis system | Edvotek Gel Electrophoresis |
Buffer | TAE |
DNA stain | SYBR Safe |
Gel images |
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Protocol notes | Taq Polymerase used: NEB OneTaq Hot Start Quick-Load 2x MM w/ Standard Buffer (product # M0488S) DNA Ladder: NEB 1kb Plus DNA Ladder for Safe Strains (product # N0559S) 3/4/25 conducted single arthropod PCR Primers: Arthropod_F (LCO1490), Arthropod_R (HCO2198) Annealing temp 49°C 3/25/25 conducted single Wolbachia PCR Primers: Wolbachia_F (16S_WspecF), Wolbachia_R (16S_WspecR) Annealing temp 55°C |
Results |
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Wolbachia presence | Yes |
Confidence level | High |
Explanation of confidence level | All controls worked. Fruit flies have often test positive for Wolbachia so results are as expected. |
Wolbachia 16S sequence | Download AB1
TGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCATCCTTAGTTACCATCAGG TAATGCTGGGGACTTTAAGGAAACTGCCAGTGATAAACTGGAGGAAGGTGGGGATGATGTCAAGTCATCATGGCCCTTAT GGAGTGGGCTACACACGTGCTACAATGGTGGCTACAATGGGCTGCAAAGTCGCGAGGCTAAGCTAATCCCTTAAAAGCCA TCTCAGTTCGGATTGTACTCTGCAACTCGAGTGCATGAAGTTGGAATCGCTAGTAATCGTGGATCAGCACGCCACGGTGA ATACGTTCTCGGGTCTTGTACACACTGCCCGTCACGCCATGGGAATTGG
BLAST at The Wolbachia Project BLAST at NCBI
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Arthropod COI sequence | Download AB1
AGTTGGAACATCTTTAAGAATTTTAATTCGAGCTGAATTAGGACATCCTGGAGCATTAATTGGAGATGATCAAATTTATA ATGTAATTGTAACTGCACATGCTTTTATTATAATTTTTTTTATAGTTATACCTATTATAATTGGTGGATTTGGAAATTGA TTAGTGCCTTTAATATTAGGTGCTCCTGATATAGCATTCCCACGAATAAATAATATAAGATTTTGACTTCTACCTCCTGC TCTTTCTTTACTATTAGTAAGTAGAATAGTTGAAAATGGAGCTGGGACAGGATGAACTGTTTATCCACCTCTATCCGCTG GAATTGCTCATGGTGGAGCTTCAGTTGATTTAGCTATTTTTTCTCTACATTTAGCAGGAATTTCTTCAATTTTAGGAGCT GTAAATTTTATTACAACTGTAATTAATATACGATCAACAGGAATTTCATTAGATCGTATACCTTTATTTGTTTGATCAGT AGTTATTACTGCTTTATTATTATTATTATCACTTCCAGTACTAGCAGGAGCTATTACTATATTATTAACAGATCGAAATT TAAATAC
BLAST at The Wolbachia Project BLAST at NCBI
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Summary | The Drosophila melanogaster was found to be postive for Wolbachia. |