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  • A A student assists Colorado Parks & Wildlife employees spawning greenback cutthroat trout at the Leadville National Fish Hatchery; B greenback cutthroat trout adults in a hatchery raceway; C tissue samples collected by students to be used for genetic analysis (images taken by S. Love Stowell)

    Cutthroat trout in Colorado: A case study connecting evolution and conservation

    Learning Objectives
    Students will be able to:
    • interpret figures such as maps, phylogenies, STRUCTURE plots, and networks for species delimitation
    • identify sources of uncertainty and disagreement in real data sets
    • propose research to address or remedy uncertainty
    • construct an evidence-based argument for the management of a rare taxon
  • Fully annotated mitochondrial genome of a lichenized fungal species (Cladonia subtenuis).  This represents a visual representation of the final project result of the lesson plan. Students will submit their annotation to NCBI (GenBank) and upon acceptance of their annotation, they typically add this publicly available resource into their resume.

    A CURE-based approach to teaching genomics using mitochondrial genomes

    Learning Objectives
    • Install the appropriate programs such as Putty and WinSCP.
    • Navigate NCBI's website including their different BLAST programs (e.g., blastn, tblastx, blastp and blastx)
    • Use command-line BLAST to identify mitochondrial contigs within a whole genome assembly
    • Filter the desired sequence (using grep) and move the assembled mitochondrial genome onto your own computer (using FTP or SCP)
    • Error-correct contigs (bwa mem, samtools tview), connect and circularize organellar contigs (extending from filtered reads)
    • Transform assembled sequences into annotated genomes
    • Orient to canonical start locations in the mitochondrial genome (cox1)
    • Identify the boundaries of all coding components of the mitochondrial genome using BLAST, including: Protein coding genes (BLASTx and tBLASTX), tRNAs (proprietary programs such as tRNAscan), rRNAs (BLASTn, Chlorobox), ORFs (NCBI's ORFFinder)
    • Deposit annotation onto genome repository (NCBI)
    • Update CV/resume to reflect bioinformatics skills learned in this lesson
  • DNA

    Using CRISPR-Cas9 to teach the fundamentals of molecular biology and experimental design

    Learning Objectives
    Module 1
    • Generate a testable hypothesis that requires a creative design of reagents based on critical reading of and review of prior research.
    • Demonstrate proficiency in using molecular cloning software to analyze, manipulate and verify DNA sequences.
    • Predict the downstream effect on the mRNA and protein after successfully inserting a DNA repair template into the genome of a cell/organism.
    • Compare and contrast the processes of DNA duplication and PCR.
    • Demonstrate the ability to design primers to amplify a nucleotide sequence.
    • Analyze and evaluate the results of DNA agarose gel electrophoresis.
    Module 2
    • Identify the key features in genomic DNA, specifically those required for CRISPR-Cas9 mediated gene edits.
    • Explain how compatible ends of DNA are used to produce recombinant DNA in a ligation reaction.
    • Explain the chemical principles behind plasmid DNA purification from bacterial cultures.
    • Devise a strategy to screen clones based on antibiotic selection and the mechanism of digestion by DNA endonucleases.
    • Predict and evaluate the results of a diagnostic digest.
    Module 3
    • Explain the chemical principles behind DNA purification using phenol-chloroform extraction and ethanol precipitation.
    • Explain the key differences between DNA duplication and transcription.
    • Demonstrate the ability to perform lab work with sterile technique.
    • Compare and contrast the results of a non-denaturing vs. denaturing agarose gel.
    • Evaluate the results of a denaturing agarose gel.
    Module 4
    • Design and implement an experiment that tests the CRISPR-Cas9 principle.
    • Predict the outcome of a successful in vitro Cas9 digest.
    Presentation of Data Post Lesson
    • Summarize important background information on gene of interest from analysis of primary literature.
    • Produce figures and figure legends that clearly indicate results.
    • Organize and construct a poster that clearly and professionally displays the important aspects of the lesson.
    • Demonstrate understanding of the lesson by presenting a poster to an audience in lay terms, mid-level terms, or at an expert level.
    • Demonstrate understanding of procedures by writing a formal materials and methods paper.