June 9, 2022

Sample information
Picture
Photos by: McKenna D.
Location
Collection date 08/06/2022
Captive / Cultivated? Wild-caught
Group Pingry School
Observations

Collected outside Pingry senior area.
Putative identification Arthropoda Hexapoda Insecta Hemiptera

Methods
Extraction kit DNeasy (Qiagen)
DNA extraction location Abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system Standard electrophoresis system
Buffer TAE
DNA stain SYBR Safe
Gel images
Protocol notes

All bands appeared in expected locations. As this was our second time completing the protocol, we ran all experiments without controls.

Results
Wolbachia presence Yes
Confidence level High
Explanation of confidence level

Our DNA gel showed strong, consistent bands and no contamination, regardless of if we diluted the sample before PCR. However, we ran the experiment without controls. Our sequence BLASTX and BLASTN matched to Wolbachia samples, supporting our findings.
Wolbachia 16S sequence Download FASTA    Download AB1
CGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCATCCTTAGTTACCATCAGGTAATGCTGGGGACTTTAAGGAGACTGCCAGTGATAAACTGGAGGAAGGTGGGGATGATGTCAAGTCATCATGGCCCTTATGGAGTGGGCTACACACGTGCTACAATGGTGGCTACAATGGGCTGCAAAGTCGCGAGGCTAAGCCAATCCCTTAAAAGCCATCTCAGTTCGGATTGTACTCTGCAACTCGAGTGCATGAAGTTGGAATCGCTAGTAATCGTGGATCAGCACGCCACGGTGAATACGTTCTCGGGTCTTGTACACACTGCCCGTCACGCCATGGGAATTGGTTTc
BLAST at The Wolbachia Project   BLAST at NCBI
Arthropod COI sequence Download FASTA    Download AB1
CAATAAGAAGAATAATTATTCGATTAGAATTAATACAACCAGGATCAATAATTAAAAATGACCAAATCTATAATACCATCGTAACATCTCATGCATTCATTATAATTTTCTTTATAGTTATACCTATTATAATTGGAGGATTTGGAAACTGATTAGTTCCTATAATAATCGGTGCACCAGACATGGCATTCCCACGAATAAATAATATAAGATTTTGATTACTACCACCATCAATTTCACTTTTAATTGCAAGATCAATCGTAGGGTCAGGATCAGGTACAGGATGAACTGTTTACCCCCCTCTCTCAGCCCAAATTGCACATTCAGGACCTTCAGTAGACCTAACTATTTTTTCACTTCACATCGCAGGTTTAAGATCAATCATAGGAGCAATCAATTTCATTTCAACAACAATAAATATACGGCCAAAAGGAATAACTATAGAAAAAATACCCCTTTTCTGTTGATCAGTCCTAATTACAGCAGTTTTACTACTAGTATCATTACCAGTACTAGCTGGAGCAATTACAATACTACTAACAGATCGAAACTTTAATACCTCATTTTTTGACCCCACGGGTGGAGGAGACCCAATTTTATATCAACACTTATTCTGATTTTTTGGT
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Hemiptera was found to be postive for Wolbachia.
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