Captive fruit fly

March 11, 2025

Sample information
Picture
Photos by: Asia L
Location
Collection date 02/18/2025
Captive / Cultivated? Captive / Cultivated
Group Berkshire Community College
Observations
Putative identification Arthropoda Insecta Diptera Drosophilidae Drosophila Drosophila melanogaster

Methods
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
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
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
Arthropod COI sequence Download AB1
AGTTGGAACATCTTTAAGAATTTTAATTCGAGCTGAATTAGGACATCCTGGAGCATTAATTGGAGATGATCAAATTTATA ATGTAATTGTAACTGCACATGCTTTTATTATAATTTTTTTTATAGTTATACCTATTATAATTGGTGGATTTGGAAATTGA TTAGTGCCTTTAATATTAGGTGCTCCTGATATAGCATTCCCACGAATAAATAATATAAGATTTTGACTTCTACCTCCTGC TCTTTCTTTACTATTAGTAAGTAGAATAGTTGAAAATGGAGCTGGGACAGGATGAACTGTTTATCCACCTCTATCCGCTG GAATTGCTCATGGTGGAGCTTCAGTTGATTTAGCTATTTTTTCTCTACATTTAGCAGGAATTTCTTCAATTTTAGGAGCT GTAAATTTTATTACAACTGTAATTAATATACGATCAACAGGAATTTCATTAGATCGTATACCTTTATTTGTTTGATCAGT AGTTATTACTGCTTTATTATTATTATTATCACTTCCAGTACTAGCAGGAGCTATTACTATATTATTAACAGATCGAAATT TAAATAC
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Drosophila melanogaster was found to be postive for Wolbachia.
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