Grasshopper Wolbachia search

November 6, 2022

Sample information
Picture
Photos by: Me (Max Bean)
Location
Collection date 05/21/2021
Captive / Cultivated? Wild-caught
Group St. Albans School
Observations

Bug in ethanol for over a year when analyzed

Legs falling off / large
Putative identification Arthropoda Hexapoda Insecta Orthoptera

Methods
Extraction kit DNeasy (Qiagen)
DNA extraction location Partial abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniPCR
Buffer TBE
DNA stain GelGreen
Gel images
Protocol notes

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

I am highly confident in my insect not having Wolbachia because of the results from the gels. In the control for arthropod positive DNA, my test also showed positive. However while the control for Wolbachia positive was positive, and my test showed negative.  Which showed that my insect was negative for Wolbachia DNA and why I am confident in that fact.
Wolbachia 16S sequence
Arthropod COI sequence Download FASTA    Download AB1
GNNTGANCTNNNNNNTAGGAACTTCCCTTAGATTATTAATTCGAGCTGAATTAGGNCAACCTGGATTTTTAATTGNANATGATCAAATTTATAATGTAATNNTAACTGCCCATGCNTTNGTTATAATTTTTTTNNNNNTAATACCAATTNNAATTGNNGGATTCGGAAATTGATTANNNCCNNNNATATTANGANNNCCNNANATANCATTCCCNCNAATAAATAANATNANATTTTNATTACTTCNNCCTTCATTANCACTNNNTTTANNANNAANNNNNNNNNANNATNNANCTGGAACTGNATGAACTGNTNANCNNNCTNNNNNNNNNNNGAATNNNNCNNGCNGNANGANNNNNANANTNNNNAATTTNNNNTTTACATTTNNNNGGNNNNNNNTNNATTTTANNNNNNNNAAATTTTATTACNNNNNNNNNNNANNNNNNNNNNNNNGNNNNNNNNNNNNNNNNNANACNNNNATNNNNNNNANNNNNNNNTATTACNNNNNNNNNNNNNNNNNNNNCNNNACNNNNNNNNNNNNNNNNNNNNNNNATN
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Orthoptera was found to be negative for Wolbachia.
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