Sample information

Crayfish was collected from a stream in the forest during the winter.

Methods

• 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

In the first gel image (a single PCR with only Wolbachia primers), the crayfish was loaded into lane 2 (sample MTC1) and shows no bands, which is consistent with a negative result for Wolbachia.

In the second gel image (a duplex PCR with both Wolbachia and arthropod primers), the crayfish was loaded into lane 2 (sample MTC2, extracted from a separate piece of tissue than MTC1) and shows only one band, which is consistent with a negative result for Wolbachia but a positive DNA extraction for arthropod DNA.