Discover the Microbes Within!

Labs and Lectures

Getting Started

Lectures, laboratory activities, and resources for each module are listed below. The numbers in parentheses refer to the date of the updated document. The Wolbachia Project protocols were designed to optimize student success in the classroom. In some cases, teachers and collaborators have provided alternate protocols to accommodate availability of classroom resources. These are provided as alternate protocols.

Download the full materials list here: Wolbachia Materials List 2017

Insect controls (Wolbachia-infected and Wolbachia-uninfected) and PCR primers are available from the Bordenstein Lab. Please contact us to request these materials.

Download the journal article “Using the Wolbachia Bacterial Symbiont to Teach Inquiry-Based Science: A High School Laboratory Series.”

Download the list of National Science Education Standards aligned with this project here: Education standards


Introductory Activities (Optional)

Teachers and collaborators have designed the following activities to accompany the project. They serve as great introductory material to the lab series.

  • Pipetting 101: This lab introduces students to the proper use and care of micropipettes.  Pipetting Lab (4/16)
  • Insects and Their Relatives: This activity introduces students to the characteristics that distinguish insects from their closest relatives and the similarities common to Arthropods.  Insects and Relatives Activity
  • Wolbachia Project Activity: This simulation of the different labs protocols can be used as an introduction for students or as an overview at professional development workshops.  Activity Description; Insect Order Guide

 

Lab 1: Insect Identification (Biodiversity)

Goal:
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)

Lecture: Insect Identification (04/17)
Lab Activity: Insect Identification (Biodiversity) (4/17)
Resource: Insect Identification Key

 

Lab 2: DNA Extraction (Molecular Biology)

Goal:
To introduce students to DNA extraction techniques and to isolate genomic DNA from insects and Wolbachia, the endosymbiotic bacteria that live within the cells of over 20% 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)

Lecture: Molecular Biology Techniques I and II: DNA and PCR (04/17)
Lab Activity: DNA Extraction (Molecular Biology) (4/17)


Alternate Protocol: DNA Extraction using Edward’s Buffer
Alternate Protocol: DNA Extraction using Sigma-Aldrich Extract-and-Amp
Alternate Protocol: DNA Extraction using Promega Wizard SV Genomic DNA Purification System
Alternate Protocol: DNA Extraction using the BioRad InstaGene Matrix Kit (#732-6030EDU); Teacher Copy; Student Copy

 

Lab 3: Polymerase Chain Reaction (Molecular Biology)

Goal:
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.

Learning Objectives:gallery2-5
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 three morphospecies and three controls 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: 50 minutes

Lecture: PCR – A Review (04/17)
Lab Activity: PCR (Molecular Biology) (4/17)
Resource: PCR_block


Alternate Protocol: PCR with ReadyToGo Beads (Old protocol) (11/16)

Note regarding the PCR protocols

 

Lab 4: Gel Electrophoresis (Molecular Biology)

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

Learning Objectives:gallery4-4
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

Lecture: Molecular Biology Techniques III: Gel Electrophoresis (08/14)
Lab Activity: Gel Electrophoresis (Molecular biology) (4/17)


Alternate Protocol: DNA Gels using Sodium Boric Acid Buffer
Alternate Protocol: Fast Blue DNA Stain
Alternate Protocol: BioRad Gel Electrophoresis classroom study kit (#166-2500EDU).

 

Lab 5: DNA Sequence Analysis (Bioinformatics)

Goals:NCBI
Module 1: To show the ways in which the NCBI online database classifies and organizes information on DNA sequences, evolutionary relationships, and nucleutide-blast-cover copyscientific publications.

Module 2: To identify an unknown nucleotide sequence from an insect endosymbiont by using the NCBI search tool BLAST

Teaching Time: 45 minutes

Lecture: Bioinformatics Lab – NCBI, BLAST, and Phylogeny (04/17)
Lab Activity: Bioinformatics (4/17)
Resources: Using BLAST to determine evolutionary relationships (04/17),
Bioinformatics Exercises, Sea Mammal Nucleotides (.txt file)

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