Tortistilus wickhami voucher

December 1, 2022

Sample information
Picture
Photos by: Chase Hartzell, Heather Ahlstrom
Location
Collection date 09/05/2022
Captive / Cultivated? Wild-caught
Group College of Eastern Idaho
Observations
  1. Tortistilus wickhami voucher – Three Cornered Alfalfa Hopper- Bright green brush leaves

Description: Bright green, small leaf like body

Body length: .76 cm. Found by Chase Hartzell

UTM Coordinates: 12T 0491965 4797196

Arthropod: + Wolbachia: +
Putative identification Arthropoda Hexapoda Insecta Hemiptera Membracidae Tortistilus Tortistilus wickhami

Methods
Extraction kit Edwards Buffer
DNA extraction location Abdomen
Single or Duplex PCR Duplex Reaction
Gel electrophoresis system Standard electrophoresis system
Buffer TAE
DNA stain GelGreen
Gel images
Protocol notes

DNA Annotated protocol in Wolbachia project Lab2: DNA Extraction Edward’s Buffer

Results
Wolbachia presence Yes
Confidence level High
Explanation of confidence level

We tested and retested our sample and then re did the protocol and did the PCR again and sent it to the Wolbachia Project Lab. It was sequenced and then confirmed on Blast as a match.
Wolbachia 16S sequence
>Wol17 GTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCATCCTTAGTTGCCATCAGGTAATGCTGAGCACTTTAAGGAAACTCCCAGTGATAAGCTGGAGGAAGATGGGGATGATGTCAAGTCATCATGGCCTTTATGAAGTGGGCTACACACGTGCTACAATGGTGTCTACAATGGGCTGCAAGGTGCGCAAGCCTAAGCTAATCCCTAAAAGACATCTCAGTTCGGATTGTACTCTGCAACTCGAGTGCATGAAGTTGGAATCGCTAGTAATCGTGGATCAGCATGCCACGGTGAATACGTTCTCGGGTCTTGTACACACTGCCCGTCACGCCATGGGAATTGGTTT
BLAST at The Wolbachia Project   BLAST at NCBI
Arthropod COI sequence
>Arthopod17_CO1F ACTTAGTGTTATTATTCGAGTTGAACTAGGGCAACCTGGACCATTACTTAATAATGATCAAATTTATAATACAATTATCACTTCTCATGCCTTCATTATAATTTTCTTCATAGTTATACCAATTATGATTGGGGGTTTTGGTAATTGATTGTTACCTATTATAATTGGGGCTCCAGATATGGCTTTTCCACGAATAAACAATATAAGATTTTGATTATTACCCCCATCTATTAGATTAATTTTAATTAGATCCATAGTAGAAACAGGGGCTGGGACGGGATGAACAGTATACCCACCATTATCAAGAAATATTGCTCACTCAGGGGCTAGAGTAGACTTTGCTATTTTTTCATTACATTTAGCTGGAATTTCATCTATTCTAGGAGCAATTAATTTCATTACAACTATAATAAACATGCGAGTACAAGGTATAAATTTAGATAAAACACCTCTATTTGTTTGATCAGTAGTTATCACTGCATTCTTACTCCTTATATCATTACCAGTTCTAGCGGGAGCAATTACCATGCTATTAGCTGACCGAAATATAAATACTTCA
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Tortistilus wickhami was found to be postive for Wolbachia.
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