Sample information |
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Picture |
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Location | |
Collection date | 09/09/2024 |
Captive / Cultivated? | Wild-caught |
Group | Bluegrass Community and Technical College |
Observations | The specimen was collected using a flying insect trap containing banana peels and vinegar per the Arthropod Collection Traps Guide provided by the Wolbachia Project. The trap was hung from a tree branch adjacent to a small creek in a suburban backyard. The specimen was identified as a Stripe-horned Green Lacewing using the iNaturalist app. |
Putative identification | Arthropoda Hexapoda Insecta Neuroptera Chrysopidae |
Methods |
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Extraction kit | DNeasy (Qiagen) blood and tissue kit |
DNA extraction location | Whole arthropod |
Single or Duplex PCR | Single Reaction |
Gel electrophoresis system | Standard electrophoresis system |
Buffer | TAE |
DNA stain | Ethidium Bromide |
Gel images |
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Protocol notes | GoTaqGreen polymerase was used. Reaction conditions were per Lab 3: PCR (Wolbachia Project Guide) for single reaction PCRs (separate reactions were run with the Wolbachia primers and the cytochrome oxidase primers). The PCR reaction volume was 25 uL. 5 uL of each reaction was run on a 2% agarose gel. The relevant lanes, lanes 8 and 9, for the specimen (#24) are boxed. |
Results |
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Wolbachia presence | No |
Confidence level | High |
Explanation of confidence level | No PCR product was obtained in the reaction using the Wolbachia primers (WO, Lane 8), while the reaction using the cytochrome oxidase 1 (CO, Lane 9) produced a ~700 bp product, which is the correct size for the CO1 product, indicating the DNA isolated from the specimen is of suitable quality for PCR. All control reactions (Lanes 10 through 15) worked as they should. The positive control is DNA extracted from a control insect infected with Wolbachia, while the negative control is DNA extracted from an uninfected insect. Both controls were provided by the Wolbachia Project. |
Wolbachia 16S sequence |
Sequencing not performed
BLAST at The Wolbachia Project BLAST at NCBI
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Arthropod COI sequence |
Sequencing not performed
BLAST at The Wolbachia Project BLAST at NCBI
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Summary | The Chrysopidae was found to be negative for Wolbachia. |