Harmonia axyridis

April 17, 2025

Sample information
Picture
Photos by: Nourah K.
Location
Collection date 11/04/2030
Captive / Cultivated? Wild-caught
Group Benedictine University
Observations

collected by Fall 2024 students
Putative identification Arthropoda Insecta Coleoptera Coccinellidae Harmonia Harmonia axyridis

Methods
Extraction kit DNeasy (Qiagen) blood and tissue kit
DNA extraction location Abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system Standard electrophoresis system
Buffer TAE
DNA stain SYBR Safe
Gel images
Protocol notes

STEPS OF ISOLATION:

•Prepare sample

•Cell lysis and DNA precipitation

•DNA Purification

DNA Elution

STEPS OF PCR:

•PCR

•Purpose/ Goal: The purpose of PCR is to amplify a specific DNA segment

•It is essential in identifying an organism and identifying infection

•Process:

•Denaturation

•DNA is heated which separates the double-stranded DNA into single strands by breaking hydrogen bonds

•Temperature: 94 – 96 °C

•Annealing

•allow primers to bind (anneal) to their complementary sequences on the single-stranded DNA

•Temperature: 49-55 °C

•Elongation

•Synthesizes new DNA strands as Taq polymerase adds nucleotides to the primers

Temperature: 72 ° C

Results
Wolbachia presence No
Confidence level Low
Explanation of confidence level

Gel electrophoresis was unsuccessful as the pipette tip was penetrated the gel, therefore I was unable to determine Wolbachia presence
Wolbachia 16S sequence
Arthropod COI sequence Download FASTA   
Summary The Harmonia axyridis was found to be negative for Wolbachia.
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