Arthropod collection (dubia roach)

April 3, 2025

Sample information
Picture
Photos by: Jake D
Location
Collection date 02/20/2025
Captive / Cultivated? Captive / Cultivated
Group Berkshire Community College
Observations

Captive bred by and ordered from dubiaroaches.com
Putative identification Arthropoda Insecta Blattodea Blaberidae Blaptica Blaptica dubia

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

Taq polymerase: New England Biolabs OneTaq hot start quick-load 2x MM w/ standard buffer (product #M0488S)

DNA ladder: New England Biolabs 1 kb plus DNA ladder safe stains (product N0559S)

Duplex PCR set up on 3/27/25 with both arthropod and wolbachia primers at an annealing temperature of 49 Celsius.

Lanes
1- DNA ladder
2- ELB1 sample from specimen 1 dubia roach
3- JBD1 sample from specimen 2 dubia roach
4- AC+ wolbachia positive fruit fly control
5- AC- wolbachia negative fruit fly control
6- DNA+ wolbachia positive arthropod DNA Control
7- water

Second gel(w/out annotation) is a single PCR set up on 3/20/25 was used but contained too much DNA, however the control samples showed expected results. Lanes contain the same samples in both gels

Results
Wolbachia presence No
Confidence level Low
Explanation of confidence level

Confidence level is low because there was very slight banding at 700 base pairs for arthropod CO1 despite smearing in lane 1 and 2, but no faint bands at 400. However all control samples show bands as expected indicating PCR was done correctly.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Blaptica dubia was found to be negative for Wolbachia.
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