Pillbug AA

Sample information
Picture	Photos by: Austin A. , Dawna W.
Location
Collection date	09/14/2022
Captive / Cultivated?	Wild-caught
Group	Woburn Memorial High School
Observations	Found it under a rock when the temperature of the local area was around 70-80 Fahrenheit.
Putative identification	Arthropoda Malacostraca Isopoda Armadillidae
Methods
Extraction kit	Edwards Buffer
DNA extraction location	Abdomen
Single or Duplex PCR	Single Reaction
Gel electrophoresis system	Edvotek Gel Electrophoresis
Buffer	TBE
DNA stain	GelGreen
Gel images
Protocol notes	Gel was moved a bit after having the DNA injected into the wells. Used a 2% agarose gel instead of a 1%. DNA ladder is in well 1, + DNA control is in well 2, water is in well 3, pillbug is in well 4, Wolbachia + is in well 5, Wolbachia – is in well 6, Ant is in well 7
Results
Wolbachia presence	Yes
Confidence level	High
Explanation of confidence level	All of the controls, positive and negative, worked. If you look very closely there is a dim band closest to where a DNA band for Wolbachia would be expected
Wolbachia 16S sequence	5'CAT ACC TAT TCG AAG GGA TAG 3' Forward, 5' AGC TTC GAG TGA AAC CAA TTC 3' Reverse BLAST at The Wolbachia Project BLAST at NCBI
Arthropod COI sequence	5' GGT CAA CAA ATC ATA AAG ATA TTG G 3' Forward, 5' TAA ACT TCA GGG TGA CCA AAA AAT CA 3' Reverse BLAST at The Wolbachia Project BLAST at NCBI
Summary	The Armadillidae was found to be postive for Wolbachia.

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The Wolbachia Project Database