T. Viridis (Long-Jawed Orbweaver)

April 1, 2025

Sample information
Picture
Photos by: Graham B-M
Location
Collection date 02/18/2025
Captive / Cultivated? Wild-caught
Group Berkshire Community College
Observations

Green Arachnid with a red dot at base of abdomen with white lines extending down abdomen.
Putative identification Arthropoda Chelicerata Arachnida Araneae Tetragnathidae Tetragnatha Tetragnatha viridis

Methods
Extraction kit Monarch DNA extraction (NEB)
DNA extraction location Whole arthropod
Single or Duplex PCR Single Reaction
Gel electrophoresis system Edvotek Gel Electrophoresis
Buffer TAE
DNA stain SYBR Safe
Gel images
Protocol notes

For DNA extraction Lysis process lasted 35 minutes with vortexing solution every 7 minutes. Used insect adaptation of New England Biolabs Monarch spin gDNA extraction protocol. Used the Taq Master mix 2x. The Polymerase we used was NEB One Taq Hot start quick load 2x MM w/ stand buffer (M0488s) we preformed two PCR’s the first one on March 4th, 2025 for Arthropod CO1 primers and the second on March 25th, 2025 for Wolbachia 16s primers. We ran both PCR results through an Edvotek electrophoresis set to 125v and 0.2 amps current, for 25 minutes. Then utilized a blue box to read the results.

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

Due to the 8 controls over the two separate PCR results all reading as the correct results gives me a high confidence level that the variables that were tested are actual results and not causation of a false positive/ false negative.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Tetragnatha viridis was found to be negative for Wolbachia.
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