Pitt-kit Equipment Loan Program
After attending a Department of Biological Sciences workshop, participants are eligible to request science kits containing all the materials, supplies, and equipment required to perform workshop protocols with their students at no cost to the teacher or school district.
Kit contents may be individually tailored for specific needs and goals as determined by discussion between the teacher and the Outreach Coordinator upon receipt of the Kit Application.
Pitt Kits Curricula
As a prerequisite to the any DNA techniques Pit Kit, we recommend our Micropipetting Practice Pit Kit. This activity teaches the skill of working with very small volumes using a micropipettor. This hands-on activity uses four colored water solutions and has students practice using four different micropipettors. This kit can be modified to include an introduction to using a vortexer and/or microcentrifuge.
The process by which bacteria take up foreign DNA is transformation. Bacteria may be transformed with plasmids containing antibiotic resistance genes (pAMP or pKAN), or the pGLO plasmid that gives ampicillin resistance along with green glowing phenotype via the green fluorescent protein gene in the presence of UV light and arabinose sugar. This protocol may be used along with the recombinant DNA protocol, or as a separate experiment.
In this protocol, DNA plasmids with ampicillin and kanamycin resistance genes are cut with restriction enzymes, then religated together with a ligase enzyme in an attempt to form a plasmid containing the resistance genes for both antibiotics. NOTE: Recombination protocol also requires Bacterial Transformation Pitt Kit materials.
DNA Isolation/Restriction Digest/Electrophoresis
In this protocol, plasmid DNA is isolated, then digested with restriction enzymes and electrophoresis in order to observe banding patterns. Alternatively, plasmid DNA can be provided for the restriction reaction, or plasmid DNA may be provided as digested and undigested samples that are ready to load onto a gel.
This activity is designed to guide students through the process of protein synthesis and processing. This is a hands‐on activity in which students will move through five stations, each of which represents an organelle within the cell. At each station, the students perform a specific task associated with proper protein formation:
- Nucleus: Transcribe DNA
- Ribosome: Translate mRNA into individual protein domains and attach these domains
- Endoplasmic Reticulum: Fold their protein
- Golgi: Add a molecular tag to sort their protein to its final destination
- Cell Membrane: Deliver the protein to the cell membrane and learn more about the specific protein they created
Are Your Foods Genetically Modified?
Advances in genetic engineering have resulted in genetically modified plants of commercial importance. These plants have been engineered to contain genes that encode herbicide resistance, insect resistance, drought tolerance, frost tolerance and other traits. Most Americans would probably be surprised to learn that more than 60% of fresh vegetables and processed foods sold in supermarkets today are genetically modified. The laboratory activity uses a rapid method for extracting DNA from plants and food products. Then polymerase chain reaction (PCR) is used to detect for evidence of the 35S promoter that drives expression of many trans-genes found in most genetically modified foods.
PCR Amplification of D1S80 Locus
This protocol may be used as part of a "crime solving" demonstration. The polymerase chain reaction amplifies DNA at the D1S80 locus---a noncoding region on chromosome 1 composed of 16 base pair repeats that can display heterozygous or homozygous phenotypes. This is one of the same analysis crime labs use to determine if a suspects DNA is the same as that found at a crime scene. In this activity each student extracts DNA from his or her own cheek cells as a template for the PCR. After PCR, electrophoresis is used and students will see that their DNA is different for each student. They will also be able to determine if they are homozygous or heterozygous for the D1S80 locus.
Bioprospecting For Biofuel Enzymes
Before petroleum was used in automobiles, the earliest vehicles were designed to run on ethanol. In today’s world as fossil fuel shortages are becoming a concern, the use of ethanol for vehicular engines is once again being sought. Like petroleum, ethanol allows the engines to work and function properly, but more importantly it can be renewed. In this module, students learn about the importance of Biofuels, how they are developed, and use a colormetric assay to search for a biological source for an enzyme required for biofuel production. This module also allows students to use a purified enzyme, spectroscopy to create a standard curve, and inquiry-driven experimental design to explore conditions that affect enzyme function.
This module introduces students to six examples of model organisms used in scientific research. By observing these organisms, students learn why we use model systems, the advantages and disadvantages of each organism, and how each model organisms fits specific research scenarios.
Sea Urchin Development
Egg activation and early cleavage of the sea urchin embryo resemble early human development. In this activity students collect gametes from live sea urchins, initiate fertilization and observe early embryogenesis under a microscope. Experiments can be conducted to see the affect various environmental factors have on sea urchin development. This lab is well suited for an inquiry-based approach to education allowing the student to be the investigator.
The Strawberry Caper
This activity harnesses middle school students’ healthy skepticism and budding independence as thinkers by immersing them in a scenario in which they are required to investigate a patent infringement claim made against an organic strawberry grower by a large commercial company. This is an exercise in critical analysis and inquiry-based thinking and allows students to analyze data, formulate relevant questions, and test/revise hypotheses. In short, this exercise forces students to think, ask questions and design experiments to proceed and can be adapted to the high school level.
Experimental techniques include:
- Phenotypic analysis
- DNA extraction
- Gel electrophoresis
Ectotherm ER: Frogs Under the Weather
This curriculum is based on the research of Dr. Richards-Zawacki, whose lab investigates how climate change and host/pathogen ecology shapes the dynamics of wildlife diseases. Students investigate possible causes for amphibian declines through an experiment that uses thermal model frogs to learn how changes in climate impact frog survival.
"Outbreak" is a simulation that uses the concept of infectious disease to allow students to analyze data, formulate relevant questions, and test/revise hypotheses. Students are presented with a scenario and provided data to analyze. They must use critical analysis and inquiry-based thinking to solve the case of a possible outbreak.
In the scenario, the causative agent is unknown. It is not known if the "disease" is contagious or due to environmental factors. Teachers and students play the part of agents representing the Center for Disease Control. It is up to the class to analyze existing facts and data and ask the pertinent questions that will allow investigation to proceed to hypothesis, hypothesis testing, and diagnosis so that the outbreak can be controlled. During the exercise, students will become acquainted with current science and technology (PCR, electron microscopy, electrophoresis, cell culture), as well as problem solving techniques.
Prior to spending the day at Pitt for this exercise, the teacher will guide students through a pre-lab activity that introduces them to the problem at hand and initiates their critical thinking. Once on-site, the students will step into the scenario. They will be greeted as CDC field agents by "Panther Hospital" chief of staff and briefed on the status of the situation. Next, the students will proceed through four laboratory rotations during which they will collect and analyze data, and interpret results in the context of the possible outbreak. Finally students will be asked to determine the cause of the illness and provide treatment recommendations.