|Captive / Cultivated?||Wild-caught|
Ant specimen was found in a terrestrial environment – decomposing leaves next to a decomposing log within The Pingry School’s forest. It was caught at the beginning of June 2022 (start of Summer/end of Spring), around noon. Temperature was 23.9 degrees celsius. Attached is a picture of the place I found the Ant in (with the ant in the picture as well).
|Putative identification||Hexapoda Insecta Hymenoptera Formicidae Camponotus Camponotus chromaiodes|
|Extraction kit||DNeasy (Qiagen)|
|DNA extraction location||Whole arthropod|
|Single or Duplex PCR||Single Reaction|
|Gel electrophoresis system||Standard electrophoresis system|
|DNA stain||SYBR Safe|
The gel contains me and my lab partner’s DNA – my Insect and Wolbachia DNA are in lanes 4 and 5 respectively.
Lane 1 – DNA Ladder
Lane 2 – Partner’s ant
Lane 3 – Partner’s wolbachia test
Lane 4 – My Ant Test
Lane 5 – My Wolbachia test
The expected fragment for both Wolbachia 16S and Arthropod CO1 both showed up, in lanes 5 and 4 respectively. Wolbachia 16S has a base pair length of 438 base pairs, and in my gel’s lane 5 there is a band under the 500 base pair DNA ladder band – indicating the expected fragment. Arthropod CO1 has a base pair length of 708 base pairs, and in lane 4, there is a band between the 1000 base pair and 500 base pair length bands – indicating the expected fragment. DNA Sequencing further supported the identification of Carpenter Ant and + Wolbachia DNA, as when I input my sequences into BLAST, we observed very strong matches for both Wolbachia and Carpenter ant DNA.
|Explanation of confidence level||
My protocols worked as expected. I had minimal problems with the protocol, and I had bands for both Insect and Wolbachia DNA in the gel. Additionally, our classroom sent our samples to be sequenced, and the sequencing that got returned further confirmed my confidence – I input my sequences into BLAST, and I got very good matches with my sequence.
|Wolbachia 16S sequence|| Download FASTA
GCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCATCCTTAG TTACCATCAGGTAATGCTGGGGACTTTAAGGAAACTGCCAGTGATAAACTGGAGGAAGGTGGGGATGATGTCAAGTCATC ATGGCCCTTATGGAGTGGGCTACACACGTGCTACAATGGTGGCTACAATGGGCTGCAAAGTCGCGAGGCTAAGCTAATCC CTTAAAAGCCATCTCAGTTCGGATTGTACTCTGCAACTCGAGTGCATGAAGTTGGAATCGCTAGTAATCGTGGATCAGCA CGCCACGGTGAATACGTTCTCGGGTCTTGTACACACTGCCCGTCACGCCATGGGAATTNBLAST at The Wolbachia Project BLAST at NCBI
|Arthropod COI sequence|| Download FASTA
TGGCTCCTCTATAAGAATAATCATTCGACTAGAGTTGGGATCTCCTGATTCACTAATTCTTAATGATC AAACTTTCAATACCATCGTTACAAGTCATGCTTTTATTATAATTTTTTTTATAGTTATACCTTTTATAATTGGGGGATTT GGTAATTTTTTAATTCCCCTTATACTAGGATCTCCTGATATAGCTTACCCCCGTTTAAATAACATAAGATTTTGATTACT TCCCCCATCGATCTCCTTATTAATCCTAAGAAATTTTATTAATGAAGGATCTGGAACTGGTTGGACTGTCTACCCCCCTC TATCATCAAATACCTTCCATAGTGGCCCCTCTATTGACCTGACTATCTTTTCTCTCCATATTGCTGGTATATCCTCAATT ATAGGAGCAATCAATTTTATTTCAACAATTATAAATATACATAATTCCAATATTTCCCTAGATAAAATTCCCTTATTAGT ATGGTCTATTCTTATTACAGCTATTCTCCTTCTTCTGTCCCTACCTGTTCTAGCAGGAGCTATTACAATACTACTAACAG ACCGAAATCTTAATACTTCATTTTTCBLAST at The Wolbachia Project BLAST at NCBI
|Summary||The Camponotus chromaiodes was found to be postive for Wolbachia.|