Wolbachia JF 2022

Sample information

Picture
Photos by: Jessica F
Location
Collection date 09/14/2021
Captive / Cultivated? Wild-caught
Group Woburn Memorial High School
Observations

Leaf hopper found in Woburn Memorial highschool grass. It is green and small.

Putative identification Arthropoda Hexapoda Insecta Hemiptera

Methods

Extraction kit Edwards Buffer
DNA extraction location Abdomen
Single or Duplex PCR Duplex Reaction
Gel electrophoresis system MiniOne
Buffer TBE
DNA stain GelGreen
Gel images
Protocol notes

Duplex PCR reaction was preformed.

Results

Wolbachia presence Unknown
Confidence level
Explanation of confidence level

In this lab the question was if our arthropod contained Wolbachia DNA. The purpose of the lab was to isolate DNA from organisms and determine if there is Wolbachia. During this lab we used PCR to amplify arthropod and Wolbachia DNA. The data shows in image 2, that no band is shown in well 2. This well contained my arthropod sample. I can not draw a conclusion from my results because neither the 708 bp band or 438 bp band was present. I think this data may be caused from not properly isolating and purifying the DNA from the sample. In image 1,  well 5 PCR #2 the wolbachia double band is present. In well 6, there is only the 708 band which means this sample is wolbachia negative. These samples worked in run 1 (image 1) but were left out in run 2 (image 2). I am unable to draw conclusions for my run 2 results. There is a band present in my DNA control and in my water control. These results make it unable to understand my results because there was contamination in my controls. 

Wolbachia 16S sequence
Primers provided by Wolbachia project
BLAST at The Wolbachia Project   BLAST at NCBI
Arthropod COI sequence
Primers provided by Wolbachia Project
BLAST at The Wolbachia Project   BLAST at NCBI
Summary
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