Crayfish of Cambaridae (Family)

February 25, 2025

Sample information
Picture
Photos by: Michael C
Location
Collection date 02/17/2025
Captive / Cultivated? Wild-caught
Group Berkshire Community College
Observations

-Upon collection Arthropod appeared
Putative identification Arthropoda Malacostraca Decapoda

Methods
Extraction kit Monarch DNA extraction (NEB)
DNA extraction location Partial abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system Edvotek Gel Electrophoresis
Buffer TAE
DNA stain SYBR Safe
Gel images
Protocol notes
  • Taq Polymerase Used – New England Biolabs OneTaq Hot Start Quick-Load 2x MM w/ Standard Buffer (product #M0488S)
  • DNA Ladder Used – New England Biolabs 1kb Plus DNA Ladder for Safe Stains (product #N0559S)

Single Arthropod PCR set up on March 6th, 2025 to amplify fragment of the CO1 cytochrome oxidase gene using Arthropod_F (LCO1490) and Arthropod_R (HCO2198) Primers with an annealing temperature of 49°C.

Separate Single Wolbachia PCR set up on March 27th, 2025 to amplify fragment of the 16S ribosomal RNA gene using Wolbachia_F (16S_WspecF) and Wolbachia_R (16S_WspecR) Primers with an annealing temperature of 55°C.

Results
Wolbachia presence No
Confidence level Medium
Explanation of confidence level
Wolbachia 16S sequence
Arthropod COI sequence Download AB1
TTCATTAAGGTTAATTATTCGAGTTGAATTAGGCCAGGTAGGTAGATTGATTGGAGACGACCAGATTTATAATGTGGTAG TAACAGCTCATGCTTTTGTTATAATTTTTTTTATAGTAATACCAATTATGATTGGAGGGTTTGGAAATTGGTTAATTCCT TTGATGCTTGGGGCTCCGGATATAGCTTTTCCTCGAATAAATAATATAAGATTTTGGTTGCTTCCTTTTTCTCTGACATT ATTATTAGCTAGGGGTATAGTAGAGAGGGGAGTGGGTACTGGGTGAACTGTTTATCCCCCTCTTGCTTCAGCAATCGCTC ATGCGGGTGCATCAGTAGATCTTGGAATTTTTTCTTTACATTTGGCTGGAGTATCCTCTATTTTAGGGTCAGTTAATTTT ATAACAACAGCTATCAATATACGGATGGTTGGTATAACTATGGATCGTATACCGTTATTTGTTTGATCTGTATTTATTAC TACTATTTTATTATTATTGTCTTTGCCTGTATTAGCAGGGGCAATC
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Decapoda was found to be negative for Wolbachia.
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