Wolbachia

Lab 1

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

Upon completion of this activity, students will:

Describe the diversity of insects collected in one location.
Identify insects using taxonomic classification.
Sort insects into “morphospecies” – similar looking species.
Prepare specimens for molecular studies.

Lab 2

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.

Upon completion of this activity, students will:

Isolate total genomic DNA from morphospecies identified in the Insect Identification Lab.
Develop pipetting skills to accurately aliquot small volumes.

Lab 3

Goal: To screen for Wolbachia pipientis 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.

Upon completion of this activity, students will:

Amplify DNA extracted from two morphospecies and three controls using PCR.
Understand the basic principles of PCR.

Lab 4

Goal: To determine the presence or absence of DNA that has been amplified through PCR and to quantify the size (length of the DNA molecule) of the product.

Upon completion of this activity, students will:

Determine the presence and size of Wolbachia 16S rDNA amplified by PCR.
Develop important skills in gel electrophoresis, a widely used technique for the preparation and analysis of DNA and proteins.

Lab 5

Goals:

Module 1: To show the ways in which the NCBI online database classifies and organizes information on DNA sequences, evolutionary relationships, and scientific publications.
Module 2: To identify an unknown nucleotide sequence from an insect endosymbiont by using the NCBI search tool BLAST.