ET1

April 23, 2026

Sample information
Picture
Entry by: Erica T
Location
Collection date 03/05/2026
Captive / Cultivated? Wild-caught
Group Auburn University at Montgomery
Observations

Found near the bank of the creek. Captured using a net and allowing it to climb from the net to a vial. Temperature was 81°F/27.2°C. Likely a field cricket.
Putative identification Arthropoda Insecta Orthoptera Gryllidae Gryllus Gryllus pennsylvanicus

Methods
Extraction kit DNeasy (Qiagen) blood and tissue kit
DNA extraction location Abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniOne
Buffer 1X TBE
DNA stain GelGreen
Gel images
Protocol notes

DNA extraction: The cricket was incubated for 15 minutes at 56°C for 15 minutes. All samples were stored in refrigerator at 4°C for 1 week (until next class) before purification. Eluted DNA was incubated for 1 hour at 65°C and stored frozen at -20°C until PCR.

PCR: MiniOne Taq polymerase used. Annealing temperature for each PCR reaction: 55°C (Wolbachia) and 49°C (Arthropod). Followed protocol, no mishaps.

Gel electrophroesis: This sample was loaded in well 6 (counting from left to right) in both the Wolbachia and arthropod gel. The band appeared brighter and wider than the control band, but similar to the DNA ladder.

 

Results
Wolbachia presence Yes
Confidence level High
Explanation of confidence level

The controls worked. There was a clear line for the positive control where the base pair for wolbachia was. This sample had a clear line there as well (Well 6).
Wolbachia 16S sequence Download FASTA    Download AB1
GTGTTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCATCCTTAGTTGCCATCAGGTAATGCTGAGTACTTTAAGGAAACTGCCAGTGATAAGCTGGAGGAAGGTGGGGATGATGTCAAGTCATCATGGCCTTTATGGAGTGGGCTACACACGTGCTACAATGGTGTCTACAATGGGTTGCAAGGTGCGCAAGCCTAAGCTAATCCCCAAAAGACATCTCAGTTCGGATTGTACTCTGCAACTCGAGTACATGAAGTTGGAATCGCTAGTAATCGTGGATCAGCATGCCACGGTGAATACGTTCTCGGGTCTTGTACACACTGCCCGTCACGCCATGGGAATTG
BLAST at The Wolbachia Project   BLAST at NCBI
Arthropod COI sequence Download FASTA    Download AB1
TAGTAGGCACATCATTAAGTATTTTAATTCGAACAGAACTAGGACAACCAGGCTATTTAATTGGAGATGATCAAACTTATAATGTTATCGTAACCGCACATGCATTTATCATAATTTTCTTTATAGTTATACCTATCATAATTGGAGGATTTGGAAATTGATTAGTACCATTAATATTGGGGGCTCCAGATATGGCCTTCCCACGAATAAACAATATAAGATTTTGACTTCTACCCCCATCATTAACCCTTTTATTAACCAGAAGAATAGTTGAAAATGGTGCAGGAACAGGATGAACAGTATACCCACCTCTATCAACAGGAATCGCTCACGCGGGAGCATCCGTCGATTTAGCTATCTTCTCACTACATTTAGCAGGAATTTCCTCAATTCTAGGGGCTGTAAATTTTATCACTACCATAATTAATATACGAGCTCCAGGAATATCACTAGACCAAACACCATTATTTGTTTGAGCAGTCGGAATTACAGCTCTTCTATTATTATTATCCTTACCAGTTTTAGCTGGTGCTATTACAATATTGCTTACAGATCGAAATTTAAATACATCATTTTTTGATCCAGCAGGAGGAGGGGATCCAATCTTATATCAACATCTATT
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Gryllus pennsylvanicus was found to be postive for Wolbachia.
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