Black carpenter ant

Sample information

Picture
Photos by: Sarah B.
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 Hexapoda Insecta Hymenoptera Formicidae Camponotus Camponotus pennsylvanicus

Methods

Extraction kit DNeasy (Qiagen)
DNA extraction location Abdomen
Gel electrophoresis system MiniOne
Buffer TBE
DNA stain GelGreen
Gel images
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

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
Arthropod COI sequence Download FASTA    Download AB1
AATTGGCTCCTCTATAAGAATAATCATTCGACTAGAGTTAGGATCTCCTGATTCACTAATTCTTAATGATCAAACTTTCAATACCATCGTTACAAGTCATGCTTTTATTATAATTTTTTTTATAGTTATACCTTTTATAATTGGGGGATTTGGTAATTTTTTAATTCCACTTATACTAGGATCTCCTGATATAGCTTACCCTCGTTTAAATAACATAAGATTTTGATTACTTCCCCCATCGATCTCCTTATTAATCCTAAGAAATTTTATTAATGAAGGATCTGGAACTGGTTGAACTATCTACCCCCCTCTATCATCAAATACCTTCCATAGTGGCCCCTCTATTGACCTGACTATCTTTTCTCTCCATATTGCTGGTATATCCTCAATTATAGGAGCAATCAATTTTATTTCAACAATTATAAATATACATAATTCCAATATTTCCCTAGATAAAATTCCCTTATTAGTATGATCTATTCTTATTACAGCTATTCTCCTTCTTCTGTCCCTACCTGTTCTAGCAGGCGCTATTACAATACTACTAACAGACCGAAATCTTAATACTTCATTTTTCGATCCCTCGGG
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Camponotus pennsylvanicus was found to be postive for Wolbachia.
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