Lady Beetle

April 3, 2025

Sample information
Picture
Photos by: Darwin C.
Location
Collection date 02/09/2025
Captive / Cultivated? Captive / Cultivated
Group Berkshire Community College
Observations

Ladybird beetle found crawing on a windowsill inside a house.
Putative identification Arthropoda

Methods
Extraction kit Monarch DNA extraction (NEB)
DNA extraction location body without wings
Single or Duplex PCR Single Reaction
Gel electrophoresis system Edvotek Gel Electrophoresis
Buffer TAE
DNA stain SYBR Safe
Gel images
Protocol notes

JE5 = Lamyctes emarginatus                     JE3 = Genus Harmonia

Taq polymerase used: New England Biolavs OneTaq Hot Start Quick-Load 2xx MM w/ Standard Buffer (Product #M0488S)

DNA Ladder used: New England Biolabs 1kb Plus DNA Ladder for Safe Stains (product #N0559S)

PCR 1- Date: 3/6/2025                                                                                 Primer: Arthopod                                                                                     Annealing Temperature: 49C

PCR 2- Date : 4/3/2025.                                                                      Primers – Wolbachia.                                                             Annealing Temperature: 55C

 

Results
Wolbachia presence No
Confidence level Medium
Explanation of confidence level

Wolbachia is pretty common in arthropods and lady beetles.

The controls did work because the controls that needed a band had bands. The confidence level is medium because the           (+) Arthropod was faint, so when doing the first PCR the bands were clearly displayed and when did the second PCR the bands appeared faintly.
Wolbachia 16S sequence
Arthropod COI sequence Download AB1
NNNTTNNTTTTGNNNTATGGGCAGGAATAGTAGGAACATCGTTAAGTATTTTAATTCGGTTAGAATTAGGAACTAGAGGA AGATTAATTGGAAACGACCAAATTTATAATATAATTGTTACAGCTCATGCTTTCATTATAATTTTCTTTATAGTAATACC TATTATAATTGGGGGTTTTGGAAATTGATTAGTTCCTTTAATAATTGGAGCTCCTGATATAGCATTTCCACGATTAAATA ACATAAGATTTTGACTTTTACCCCCTGCTTTAACTCTTTTAATTTTAAGAACAATCGTAGAAATAGGGGCAGGAACAGGA TGAACTGTTTACCCTCCTCTTTCTTCTAATTTAACACATAATGGGCCTTCAGTAGATTTAGTGATTTTTAGTTTACATTT AGCAGGAATTTCCTCAATTTTAGGTGCAGTAAATTTCATTTCAACTATTATAAATATACGTCCATTTGGTATAATACTTG ATAAAACTCCTTTATTTGTATGATCTGTTCTTATTACAGCAATTCTTTTATTACTATCACTACCAGTTCTTGCAGGAGCA ATTACTATACTATTAACTGACCGAAACTTAAATTCTTCTTTTTTTGACNNAACNNGTGGGGGAGACCCAATTTTATACCA ACATTTATTTTGATTTTTTGNTNNNNNNNNANNTTAAA
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Arthropoda was found to be negative for Wolbachia.
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