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Grant Writing as a Teaching Tool in the Undergraduate Genetics Laboratory David P. Aiello Department of Biology Mercer University Genetics lab education: a vertical approach • Introductory Biology – basic lab technique – process and presentation of science – population, molecular, transmission genetics • Genetics – applications of genetics – introduction to advanced lab exercises – independent thought and critique – incorporation of primary literature – build presentation skills • Molecular Genetics – in-depth exploration of subdiscipline – careful critiques of primary literature – advanced laboratory exercises – build presentation skills – development of the undergraduate “scientist” The idea • incorporate all of the above... • fun! • intellectually stimulating to both students AND faculty • First, a phone call… – Dr. Christi Magrath, Troy University – lab proposals from her students • Grant proposal! – KO a yeast gene – several “advanced” techniques • • • • • PCR agarose gel electrophoresis DNA purification transformation phenotype analysis – process of science – independent thought – data analysis and presentation Format of the lab experience • Week 1 – introduction to yeast – project introduction • Week 2 – grant workshop • Week 3 – grants due; peer reviews due prior to next lab mtg. • Weeks 4-9 – set-up; data collection! • Week 10 – oral presentations and/or formal write-up Week 1: Introductions • • • • Why yeast? Nomenclature Life cycle Yeast lab technologies – knockouts Grant Proposal • Abstract – brief outline of goals/significance of project • Introduction – introduce topic/review of literature/relevance • Experimental design – how will you do the experiments? what steps? – necessary reagents? • Timeline – what do you expect to accomplish each week? • Expected results What should we do??? • nuclear encoded genes only • viable null mutant • assayable phenotype (examples…) – – – – – – ion tolerance (Na, Ca, Cu) pH stress osmotic stress metabolic defects growth phenotypes colony morphology • Resources: – http://www.yeastgenome.org/ – http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=P ubMed Week 3: Grants and peer review Week 3: Grants and peer review Week 4-9: Set-up and data collection • Winning proposals – mlf3∆: MCS of leflunomide sensitivity • immunosuppressant drug, inhibits G1 progression • mlf3∆ more sensitive; heat shock resistance following leflunomide treatment – sky1∆: S/T kinase; cation homeostasis • cisplatin resistance • followed Li+, Na+, Mn2+ phenotypes; extended to Mg2+ and Ca2+ – rad27∆: 5’-3’ exonuclease for long patch base excision repair; Okazaki fragment processing • slow growth and increased cell size phenotypes • UV sensitivity • increased recombination rates Week 4-9: Set-up and data collection • Week 4: – primer for KO ordered prior to start of Wk 4 – PCR of fragment; run gel • Week 5: – LiOAc transformation – pick and streak for isolation • Week 6: – screen isolated colonies (colony PCR or DNA isolation from spheroplasts) • Weeks 7-9: – students run proposed experiments Week 10: student assessment • oral presentations • formal lab write-ups? • weekly progress reports? Program assessment • student feedback – overwhelmingly positive – some frustration (but that’s good!) • advantages and disadvantages – yeast centric/in-depth exposure to model system – traditional lab exercises lacking/incorporating many into an overall project – adaptable to wide range of disciplines – time involved • mission accomplished? – fun-yes! – intellectually stimulating-yes! – vertical genetics education? (mol genetics 08S) Acknowledgments • Christi Magrath – Associate Professor, Department of Biological and Environmental Sciences, Troy University • Mercer University Department of Biology • Pam Hanson – Associate Professor of Biology, Birmingham-Southern College