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Experimental Design. Dr MP Seed m.p.seed@qmul.ac.uk. Objectives. To be able to design an experiment taking into account: Suitable controls Suitable comparisons Suitable statistics Error reduction Power If animals are used, benefit/suffering Final Data Presentation.
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Experimental Design Dr MP Seed m.p.seed@qmul.ac.uk
Objectives • To be able to design an experiment taking into account: • Suitable controls • Suitable comparisons • Suitable statistics • Error reduction • Power • If animals are used, benefit/suffering • Final Data Presentation
Science is a process of discovery through hypothesis testing
Formulation of Hypothesis or objective Experimental Unit Control Variability Control of bias Treatment choice (independent variables) End Point choice (dependent variable) Choice of design Sample size Statistical analysis planning Pilot study Protocols and SOPs (Standard Operating Procedures) Statistical Analysis Interpretation Data Presentation Choice of ‘model’
Common Errors: Lack of design – ‘Ad Hoc’ approach Historical controls Clue: variable numbers Factorial Design unbalanced Experiment size ‘Blocking’ Genetic heterogeneity (Inbred/outbred animals strains) Statistical analysis errors Assay errors
Statistics – Parametric: The Bloody Obvious Test (t-Test) ANOVA Followed by Post Hoc test Bonferroni (No more than 4 comparisons unless corrected Neuman Keuls test (does not give confidence limits) Tuley Kramer (Gives confidence Limits) Comparison of groups to one control- Dunnett’s Test
T-test Compares two groups Reduced reliability with multiple comparisons Bonferroni also, use correction factor or do not use for more than 4 comparisons.
ANOVA Post Hoc Testing: Bonferroni, Newman Keul’s, Tukey Kramer Parametric
Non Parametric ANOVA: • Kruskall Wallace (NOT Neuman Keuls as mentoned in lecture!!) • Followed by Post Hoc test eg Dunn’s test
Power analysis Effect size 10-20% Standard deviation Significance level p<0.05 Power level 80-90% Sample size unknown? Alternative hypothesis 1 / 2 tailed?
Useful Packages: Graphpad Prizm (Desktop) Minitab (Desktop) SPS Contain useful guides
Handout: Basic Principles of the Design of Animal Experiments. Festing et al, The Design of Animal Experiments, Laboratory Anumal Handbooks No14 Chapter 1.