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Biochemistry And Molecular Biology
Using QIIME to Interpret Environmental Microbial Communities in an Upper Level Metagenomics CourseLearning ObjectivesStudents will be able to:
- list and perform the steps of sequence processing and taxonomic inference.
- interpret microbial community diversity from metagenomic sequence datasets.
- compare microbial diversity within and between samples or treatments.
Tackling "Big Data" with Biology Undergrads: A Simple RNA-seq Data Analysis Tutorial Using GalaxyLearning Objectives
- Students will locate and download high-throughput sequence data and genome annotation files from publically available data repositories.
- Students will use Galaxy to create an automated computational workflow that performs sequence quality assessment, trimming, and mapping of RNA-seq data.
- Students will analyze and interpret the outputs of RNA-seq analysis programs.
- Students will identify a group of genes that is differentially expressed between treatment and control samples, and interpret the biological significance of this list of differentially expressed genes.
An undergraduate bioinformatics curriculum that teaches eukaryotic gene structureLearning ObjectivesModule 1
- Demonstrate basic skills in using the UCSC Genome Browser to navigate to a genomic region and to control the display settings for different evidence tracks.
- Explain the relationships among DNA, pre-mRNA, mRNA, and protein.
- Describe how a primary transcript (pre-mRNA) can be synthesized using a DNA molecule as the template.
- Explain the importance of the 5' and 3' regions of the gene for initiation and termination of transcription by RNA polymerase II.
- Identify the beginning and the end of a transcript using the capabilities of the genome browser.
- Explain how the primary transcript generated by RNA polymerase II is processed to become a mature mRNA, using the sequence signals identified in Module 2.
- Use the genome browser to analyze the relationships among:
- 5' capping
- 3' polyadenylation
- Identify splice donor and acceptor sites that are best supported by RNA-Seq data and TopHat splice junction predictions.
- Utilize the canonical splice donor and splice acceptor sequences to identify intron-exon boundaries.
- Determine the codons for specific amino acids and identify reading frames by examining the Base Position track in the genome browser.
- Assemble exons to maintain the open reading frame (ORF) for a given gene.
- Define the phases of the splice donor and acceptor sites and describe how they impact the maintenance of the ORF.
- Identify the start and stop codons of an assembled ORF.
- Demonstrate how alternative splicing of a gene can lead to different mRNAs.
- Show how alternative splicing can lead to the production of different polypeptides and result in drastic changes in phenotype.
Serotonin in the Pocket: Non-covalent interactions and neurotransmitter bindingLearning Objectives
- Students will design a binding site for the neurotransmitter serotonin.
- Students will be able to determine the effect of a change in molecular orientation on the affinity of the molecule for the binding site.
- Students will be able to determine the effect of a change in molecular charge on the affinity of the molecule for the binding site.
- Students will be able to better differentiate between hydrogen bond donors and acceptors.
- Students can use this knowledge to design binding sites for other metabolites.
A Close-Up Look at PCRLearning ObjectivesAt the end of this lesson students will be able to...
- Describe the role of a primer in PCR
- Predict sequence and length of PCR product based on primer sequences
- Recognize that primers are incorporated into the final PCR products and explain why
- Identify covalent and hydrogen bonds formed and broken during PCR
- Predict the structure of PCR products after each cycle of the reaction
- Explain why amplification proceeds exponentially
Evaluating the Quick Fix: Weight Loss Drugs and Cellular RespirationLearning Objectives
- Students will be able to explain how the energy from sugars is transformed into ATP via cellular respiration.
- Students will be able to predict an outcome if there is a perturbation in the cellular respiration pathway.
- Students will be able to state and evaluate a hypothesis.
- Students will be able to interpret data from a graph, and use that data to make inferences about the action of a drug.
Follow the Sulfur: Using Yeast Mutants to Study a Metabolic PathwayLearning ObjectivesAt the end of this lesson, students will be able to:
- use spot plating techniques to compare the growth of yeast strains on solid culture media.
- predict the ability of specific met deletion strains to grow on media containing various sulfur sources.
- predict how mutations in specific genes will affect the concentrations of metabolites in the pathways involved in methionine biosynthesis.
Understanding Protein Domains: A Modular ApproachLearning Objectives
- Students will be able to compare protein sequences and identify conserved regions and putative domains.
- Students will be able to obtain, examine, and compare structural models of protein domains.
- Students will be able to interpret data on protein interactions (in vitro pull-down and in vitro and in vivo functional assays)
- Students will be able to propose experiments to test protein interactions.
Fly Exercise: A Simple Experiment to Test the Physiological Effects of Exercise on a Model OrganismLearning ObjectivesStudents will:
- demonstrate understanding of the concept and details of experimental design.
- perform an organic lipid extraction to determine total lipid content.
- quantify enzyme activity, as well as triglyceride, glucose, and glycogen concentrations.
- organize their collected data into spreadsheets for statistical analyses.
- interpret the results to gain insight on the varying effects exercise has on an organism's physiology.
- graphically present their results so that trends can be easily identified.
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.
CURE-all: Large Scale Implementation of Authentic DNA Barcoding Research into First-Year Biology CurriculumLearning ObjectivesStudents will be able to: Week 1-4: Fundamentals of Science and Biology
- List the major processes involved in scientific discovery
- List the different types of scientific studies and which types can establish causation
- Design experiments with appropriate controls
- Create and evaluate phylogenetic trees
- Define taxonomy and phylogeny and explain their relationship to each other
- Explain DNA sequence divergence and how it applies to evolutionary relationships and DNA barcoding
- Define and measure biodiversity and explain its importance
- Catalog organisms using the morphospecies concept
- Geographically map organisms using smartphones and an online mapping program
- Calculate metrics of species diversity using spreadsheet software
- Use spreadsheet software to quantify and graph biodiversity at forest edges vs. interiors
- Write a formal lab report
- Extract, amplify, visualize and sequence DNA using standard molecular techniques (PCR, gel electrophoresis, Sanger sequencing)
- Explain how DNA extraction, PCR, gel electrophoresis, and Sanger sequencing work at the molecular level
- Trim and assemble raw DNA sequence data
- Taxonomically identify DNA sequences isolated from unknown organisms using BLAST
- Visualize sequence data relationships using sequence alignments and gene-based phylogenetic trees
- Map and report data in a publicly available online database
- Share data in a formal scientific poster
Investigating Cell Signaling with Gene Expression DatasetsLearning ObjectivesStudents will be able to:
- Explain the hierarchical organization of signal transduction pathways.
- Explain the role of enzymes in signal propagation and amplification.
- Recognize the centrality of signaling pathways in cellular processes, such as metabolism, cell division, or cell motility.
- Rationalize the etiologic basis of disease in terms of deranged signaling pathways.
- Use software to analyze and interpret gene expression data.
- Use an appropriate statistical method for hypotheses testing.
- Produce reports that are written in scientific style.
Taking the Hassle out of HasselbalchLearning ObjectivesStudents will be able to:
- Characterize an aqueous environment as acidic or basic.
- Explain that pKa is a measure of how easy it is to remove a proton from a molecule.
- Predict ionization state of a molecule at a particular pH based on its pKa (qualitative use of the Henderson-Hasselbalch equation).
- Calculate the ratio of protonated/unprotonated forms of ionizable groups depending on chemical characteristics and /or environment pH (quantitative use of the Henderson-Hasselbalch equation).
- Apply this knowledge in a medical context.
Lights, Camera, Acting Transport! Using role-play to teach membrane transportLearning ObjectivesAt the end of this activity, students should be able to:
- Compare and contrast the mechanisms of simple diffusion, facilitated diffusion, and active transport (both primary and secondary).
- Identify, and provide a rationale for, the mechanism(s) by which various substances cross the plasma membrane.
- Describe the steps involved in the transport of ions by the Na+/K+ pump, and explain the importance of electrogenic pumps to the generation and maintenance of membrane potentials.
- Explain the function of electrochemical gradients as potential energy sources specifically used in secondary active transport.
- Relate each molecule or ion transported by the Na+/glucose cotransporter (SGLT1) to its own concentration or electrochemical gradient, and describe which molecules travel with and against these gradients.
Using CRISPR-Cas9 to teach the fundamentals of molecular biology and experimental designLearning ObjectivesModule 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.
- 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.
- 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.
- Design and implement an experiment that tests the CRISPR-Cas9 principle.
- Predict the outcome of a successful in vitro Cas9 digest.
- 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.
The Avocado Lab: An Inquiry-Driven Exploration of an Enzymatic Browning ReactionLearning ObjectivesStudents will be able to:
- develop a testable research question and supportive hypothesis regarding the browning of damaged avocado flesh caused by the activity of avocado polyphenol oxidase (aPPO).
- design and execute a well-controlled experiment to test aPPO hypotheses.
- evaluate qualitative enzyme activity data.
- create a figure and legend to present qualitative data that tests multiple hypotheses and variables.
- search for and correctly cite primary literature to support or refute hypotheses.
- know the role of reducing reagents, pH, chelators, and temperature in reactions catalyzed by aPPO.
- explain why the effects of salt and detergent differ for aPPO experiments conducted in situ
- (in mashed avocado flesh) as compared to in vitro (on purified protein).
- discuss how substrate and cofactor availability affect aPPO reactions.
- describe how endogenous subcellular organization restricts aPPO reactions in a healthy avocado.
- evaluate food handling practices for fruits expressing PPO.
Sequence Similarity: An inquiry based and "under the hood" approach for incorporating molecular sequence...Learning ObjectivesAt the end of this lesson, students will be able to:
- Define similarity in a non-biological and biological sense when provided with two strings of letters.
- Quantify the similarity between two gene/protein sequences.
- Explain how a substitution matrix is used to quantify similarity.
- Calculate amino acid similarity scores using a scoring matrix.
- Demonstrate how to access genomic data (e.g., from NCBI nucleotide and protein databases).
- Demonstrate how to use bioinformatics tools to analyze genomic data (e.g., BLASTP), explain a simplified BLAST search algorithm including how similarity is used to perform a BLAST search, and how to evaluate the results of a BLAST search.
- Create a nearest-neighbor distance matrix.
- Create a multiple sequence alignment using a nearest-neighbor distance matrix and a phylogram based on similarity of amino acid sequences.
- Use appropriate bioinformatics sequence alignment tools to investigate a biological question.