Brown Marmorated Stink Bug Wolbachia Project

April 17, 2024

Sample information
Picture
Photos by: Gregory B
Location
Collection date 03/16/2022
Captive / Cultivated? Wild-caught
Group River Valley Community College
Observations

Found indoors. Wings/Shell removed before DNA extraction.
Putative identification Arthropoda Hexapoda Insecta Hemiptera

Methods
Extraction kit DNeasy (Qiagen) blood and tissue kit
DNA extraction location Abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniOne
Buffer TAE
DNA stain GelRed
Gel images
Protocol notes

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

I am very confident in this sample not having Wolbachia present. Our controls for detecting the amplification of Wolbachia DNA all worked well, there was zero evidence of an issue during the extraction step either. The only controls that had problems were our controls for the Arthropod CO1 DNA that was most likely hindered during the PCR prep phase. Most likely theory is poor mixing between the DNA and the primer used to amplify the CO1 sequence in our controls.
Wolbachia 16S sequence
Arthropod COI sequence Download AB1
ACAAATTACTTTATCTTCGGATATGATCTGGAATAGTAGGATCAGCTATAAGATTAATTATCCGTATCGAATTAGGACAGCCTGGAAGATTTATTGGTAATGATCAAATTTATAATGTAATTGTAACAGCACATGCATTTGTAATAATTTTCTTTATAGTAATACCAATCATAATTGGAGGATTCGGTAATTGATTAGTCCCTTTAATAATTGGAGCACCTGATATAGCCTTCCCACGATTAAATAATATAAGATTCTGATTATTACCCCCTTCATTAACTTTATTATTAATAAGAAGAATAGCAGAATCAGGAGCTGGGACAGGATGAACAGTCTATCCCCCCTTATCAAGTAATATCTCACATAGAGGAGCATCAGTTGATTTAGCTATTTTTTCCTTACATTTGGCTGGGGTTTCATCAATTTTAGGGGCAGTAAATTTTATTTCAACTATTATAAATATACGACCAACAGGTATAACCCCTGAACGAATCCCATTGTTTGTGTGATCAGTAGGAATTACTGCACTTCTTCTGTTATTATCCTTACCTGTGTTAGCAGGTGCTATTACTATACTATTAACTGACCGAAATTTTAATACCTCTTTTTTTGACCCATCAGGAGGAGGAGACCCTATTCTTTATCAACATTTATTTTGATTTTTTGGTCACCCTGGAAAGTTTAA
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Hemiptera was found to be negative for Wolbachia.
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