1. Grant Writing as a Teaching Tool in the Undergraduate Genetics Laboratory David P. Aiello Department of Biology Mercer University

2. 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”

3. 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

4. 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

5. Week 1: Introductions Why yeast? Nomenclature Life cycle Yeast lab technologies –knockouts

6. 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

7. 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.yeastgenome.org/ –http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=P ubMedhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=P ubMed

8. Week 3: Grants and peer review

10. Week 4-9: Set-up and data collection Winning proposals –mlf3∆: MCS of leflunomide sensitivity immunosuppressant drug, inhibits G 1 progression mlf3∆ more sensitive; heat shock resistance following leflunomide treatment –sky1∆: S/T kinase; cation homeostasis cisplatin resistance followed Li +, Na +, Mn 2+ phenotypes; extended to Mg 2+ and Ca 2+ –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

11. 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

12. Week 10: student assessment oral presentations formal lab write-ups? weekly progress reports?

13. 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)

14. 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