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Project Guides

The Wolbachia Project is organized as a five-part lab series. Each Project Guide has been designed to facilitate inquiry-based research experiences and optimize student success in the classroom.


Introductory Activity (Optional)

Wolbachia and Reproductive Parasitism

This activity uses manipulatives to explore the associations between Wolbachia and arthropod hosts. As a result, students will unravel the mystery of this host-microbe symbiosis and investigate its relevance to human health.


Lab 1: Insect Identification (Biodiversity)

To introduce students to the spectacular diversity and abundance of insects and to prepare specimens for DNA analysis.

Learning Objectives:gallery1-7
Upon completion of this activity, students will conduct field work to collect insects from local fauna, appreciate the ubiquity of symbiotic microbes in animals, understand how to use a taxonomic key to identify insects to Order, sort insects into “morphospecies” – similar looking species, and prepare lab notes and specimens for molecular studies.

Teaching Time: 60-90 minutes (2 class periods)

Lab Activity: Arthropod Identification (Biodiversity) (9/20)

Data Submission Form: Student Observation Form (9/20)


Lab 2: DNA Extraction (Molecular Biology)

To introduce students to DNA extraction techniques and to isolate genomic DNA from arthropods and Wolbachia, the endosymbiotic bacteria that live within the cells of about 40% of insect species.

Learning Objectives:
Upon completion of this activity, students will transition from field work and morphological classification (Lab 1) to molecular biology and biotechnology, learn about DNA as a diagnostic tool to discover unseen microbes, increase abilities in biotechnology, and understand the process of inquiry and discovery-based research. They will isolate total genomic DNA from morphospecies identified in the Insect Identification Lab.

Prerequisite Skills: Prior practice with micropipettors.

Teaching Time: 90 minutes (2 class periods – optional 45min stopping point on page 7)

Qiagen DNeasy Blood & Tissue Kit (recommended)

Edward’s Buffer

Promega Wizard SV genomic DNA purification system


Lab 3: Polymerase Chain Reaction (Molecular Biology)

To screen for Wolbachia symbiont DNA in the extracted DNA from insects using one of the most widely used biotechnology techniques in biological research, the Polymerase Chain Reaction (PCR). PCR amplifies DNA millions of times in just a few hours, so that the DNA becomes easy to detect and study in any fashion.

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Upon completion of this activity, students will use and understand one of the most useful biotechnology tools in the life sciences (PCR), understand DNA as the hereditary basis of life, utilize DNA as a diagnostic tool to discover microbes, and seamlessly transition their discovery-based science from organisms to molecules during this lab. Students will amplify DNA extracted from both morphospecies and control arthropods using Polymerase Chain Reaction (PCR). The piece of DNA used for identifying Wolbachia is the region that codes for a small subunit of the ribosomal RNA. We will refer to this piece as 16S rDNA.

Prerequisite Skills: Prior practice with micropipettors.

Teaching Time: 45 minutes

Lab Activity: PCR (Molecular Biology) 2021 (2/21)

PCR Calculator: PCR template using MiniOne Taq Master Mix (7/19)

PCR Calculator: PCR template using Promega GoTaq Green Master Mix (7/19)

Note regarding the PCR protocol


Lab 4: Gel Electrophoresis (Molecular Biology)

To determine the presence or absence of PCR products and quantify the size (length of the DNA molecule) of the product.

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Upon completion of this activity, students will have integrated scientific discovery, inquiry and biotechnology. Students will understand that DNA contains hereditary information in the form of genes, how DNA samples separate-based upon different sizes, learn how to stain and visualize DNA samples. We will be using agarose gel electrophoresis to determine the presence and size of Wolbachia 16S rDNA amplified by our PCR.

Teaching Time: 90 minutes or two class periods of 45min each

MiniOne System

Standard Gel Electrophoresis


Lab 5: Bioinformatics

Bioinformatics I: To analyze and interpret the quality of Sanger sequences, and generate a consensus DNA sequence for bioinformatics analyses.NCBI

Bioinformatics II: (1) To show the ways in which the NCBI online database classifies and nucleutide-blast-cover copyorganizes information on DNA sequences, evolutionary relationships, and scientific publications. (2) To identify an unknown nucleotide sequence from an insect endosymbiont using the NCBI search tool BLAST.

Teaching Time: 90 minutes (one class period for each activity)

I. Sanger Sequence Analysis

II. NCBI Taxonomy & BLAST Searching

Required resources: DNA Sequence Files