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The Science Behind the ACTN3 PolymorphismLearning ObjectivesThis article accompanies the lesson "The ACTN3 Polymorphism: Applications in Genetics and Physiology Teaching Laboratories." Learning objectives for the lesson include:
- Test hypotheses related to the role of ACTN3 in skeletal muscle function.
- Explain how polymorphic variants of the ACTN3 gene affect protein structure and function.
- List and explain the differences between fast twitch and slow twitch muscle fibers.
- List and explain possible roles of the ACTN3 protein in skeletal muscle function.
- Find and analyze relevant scientific publications about the relationship between ACTN3 genotype and muscle function.
- Formulate hypotheses related to the relationship between ACTN3 genotype and skeletal muscle function.
- Design experiments to test hypotheses about the role of ACTN3 in skeletal muscle function.
- Statistically analyze experimental results using relevant software.
- Present experimental results in writing.
A Short Laboratory Module to Help Infuse Metacognition during an Introductory Course-based Research ExperienceLearning Objectives
- Students will be able to evaluate the strengths and weaknesses of data.
- Students will be able to employ prior knowledge in formulating a biological research question or hypothesis.
- Students will be able to distinguish a research question from a testable hypothesis.
- Students will recognize that the following are essential elements in experimental design: identifying gaps in prior knowledge, picking an appropriate approach (ex. experimental tools and controls) for testing a hypothesis, and reproducibility and repeatability.
- Students will be able to identify appropriate experimental tools, approaches and controls to use in testing a hypothesis.
- Students will be able to accurately explain why an experimental approach they have selected is a good choice for testing a particular hypothesis.
- Students will be able to discuss whether experimental outcomes support or fail to support a particular hypothesis, and in the case of the latter, discuss possible reasons why.
Dynamic Daphnia: An inquiry-based research experience in ecology that teaches the scientific process to first-year...Learning ObjectivesStudents will be able to:
- Construct written predictions about 1 factor experiments.
- Interpret simple (2 variables) figures.
- Construct simple (2 variables) figures from data.
- Design simple 1 factor experiments with appropriate controls.
- Demonstrate proper use of standard laboratory items, including a two-stop pipette, stereomicroscope, and laboratory notebook.
- Calculate means and standard deviations.
- Given some scaffolding (instructions), select the correct statistical test for a data set, be able to run a t-test, ANOVA, chi-squared test, and linear regression in Microsoft Excel, and be able to correctly interpret their results.
- Construct and present a scientific poster.
CRISPR/Cas9 in yeast: a multi-week laboratory exercise for undergraduate studentsLearning ObjectivesWeek 1: CRISPR design
- Locate the coding sequence, flanking sequence, protein product, and characteristics of a given gene from the Saccharomyces Genome Database (https://www.yeastgenome.org/).
- Design and defend the design of guide RNA and single stranded template for DNA repair in CRISPR/Cas9 gene editing studies to generate Saccharomyces cerevisiae auxotrophic mutants.
- Describe the qualities of the vector, pML104, that allow replication and selection in bacteria and yeast as well as allow expression of necessary factors in CRISPR/Cas9 genome editing, including Cas9 and sgRNA.
- Describe the rationale of and perform procedures necessary for cloning a small cassette (i.e., sgRNA gene) into a vector (i.e., pML104) including; restriction digest, annealing of DNA strands, removal of 5’ phosphates, ligation, and transformation.
- Recognize and design appropriate controls for cloning procedures such as ligation and transformation.
- Describe the method of polymerase chain reaction (PCR), including the rationale for essential components of a reaction mixture and thermal-cycling conditions.
- Locate the binding sites of and design primers for PCR, then report the expected size of the amplification product.
- Describe and perform isolation of plasmid DNA from E. coli.
- Describe the rationale for and perform procedures to transform yeast, including the essential components of a transformation mixture and conditions necessary for transformation.
- Describe the basic conditions required for cultivating yeast.
- Describe the rationale for and perform agarose gel electrophoresis of a given size of DNA.
- Analyze DNA separated by agarose gel electrophoresis, including size estimation.
- Recognize and describe the qualities of a template for DNA repair that allows efficient DNA repair.
- Design an experiment to determine auxotrophic phenotypes.
- Predict the outcome of multi-step experiments.
- Recognize and describe conditions necessary for growth of E. coli and S. cerevisiae.
- Qualitatively and quantitatively analyze scientific data from scientific experiments, including bacterial and yeast transformation, agarose gel electrophoresis, extraction of plasmid DNA from bacteria, PCR, and auxotroph phenotypic analysis.
- Communicate science to peers through maintenance of a laboratory notebook, verbal communication with group members, and writing of a formal laboratory report written in a format acceptable for journal publication.
- Troubleshoot scientific protocols by identifying procedures that are prone to error, comparing recommended protocols to actual procedure, and using positive and negative controls to narrow the location of a potential error.
- Communicate specific potential or actual uses of CRISPR/Cas9 in science and/or medicine.
- Use various bioinformatics approaches to analyze macromolecular primary sequence and structure.
- Illustrate how DNA is replicated and genes are transmitted from one generation to the next in multiple types of organisms including bacteria, eukaryotes, viruses, and retroviruses.
- Define what a genome consists of and how the information in various genes and other sequence classes within each genome are used to store and express genetic information.
- Explain the meaning of ploidy (haploid, diploid, aneuploid etc.) and how it relates to the number of homologues of each chromosome.
- Predict the effects of mutations on the activity, structure, or stability of a protein and design appropriate experiments to assess the effects of mutations.
- Predict the growth behavior of microbes based on their growth conditions, e.g., temperature, available nutrient, aeration level, etc.
- Discuss the benefits of specific tools of modern biotechnology that are derived from naturally occurring microbes (e.g. cloning vectors, restriction enzymes, Taq polymerase, etc.)
- Accurately prepare and use reagents and perform experiments.
- When presented with an observation, develop a testable and falsifiable hypothesis.
- When provided with a hypothesis, identify the appropriate experimental observations and controllable variables.
Using Synthetic Biology and pClone Red for Authentic Research on Promoter Function: Genetics (analyzing mutant...Learning Objectives
- Describe how cells can produce proteins at the right time and correct amount.
- Diagram a bacterial promoter with −35 and −10 elements and the transcription start site.
- Describe how mutational analysis can be used to study promoter sequence requirements.
- Develop a promoter mutation hypothesis and design an experiment to test it.
- Successfully and safely manipulate DNA and Escherichia coli for ligation and transformation experiments.
- Design an experiment to verify a mutated promoter has been cloned into a destination vector.
- Design an experiment to measure the strength of a promoter.
- Analyze data showing reporter protein produced and use the data to assess promoter strength.
- Define type IIs restriction enzymes.
- Distinguish between type II and type IIs restriction enzymes.
- Explain how Golden Gate Assembly (GGA) works.
- Measure the relative strength of a promoter compared to a standard promoter.
Linking Genotype to Phenotype: The Effect of a Mutation in Gibberellic Acid Production on Plant GerminationLearning ObjectivesStudents will be able to:
- identify when germination occurs.
- score germination in the presence and absence of GA to construct graphs of collated class data of wild-type and mutant specimens.
- identify the genotype of an unknown sample based on the analysis of their graphical data.
- organize data and perform quantitative data analysis.
- explain the importance of GA for plant germination.
- connect the inheritance of a mutation with the observed phenotype.