• Three-level designs• Polynomial structured model• Non-linear optimization algorithm to find out where the optimumis likely to be located

• Verification of the predicted model by experiment

Optimization designs

Response surface methodology: BoxResponse surface methodology: Box--Behnken DesignBehnken Design

Experimental resultsExperimental results

How to do the analysis on PC ??How to do the analysis on PC ??

YPCR yield = 189.3+2.5*primer+26.44*cycles+18*Taq+5.2*Mg+3.9*Annealing-5.1*Extension-0.25*primer*cycles-17.84*primers*Taq-10.14*cycles*Taq-10.58*primers*Mg+7.8*cycles*Mg-24.85*Taq*Mg+8.4*primers*Extension-11.96*cycles*Annealing+9.4*Taq*Annealing-3.14*Mg*Annealing+0.48*primers*Extension+0.05*cycles*Extension-13.4*Taq*Extension-20.64*Mg*Extension-8.7*Annealing*Extension-0.18*primers*primers-7.18*cycles*cycles-10.77*Taq*Taq-29.9*Mg*Mg+5.86*Anealing*Anealing-28*Extension*Extension

يتوفر لدي وحدة الخدمات المرآزية بالمدينة يتوفر لدي وحدة الخدمات المرآزية بالمدينة 100100 bpbp DNA Marker (100DNA Marker (100--2500bp)2500bp)

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Agarose gel electrophoresis (2%) with 1-5 μl of the DNA marker

Commercial software available free on WWWCommercial software available free on WWW

httphttp://://wwwwww..geocitiesgeocities..comcom//SiliconValleySiliconValley//NetworkNetwork//10321032//

Applications of statistical experimental designsApplications of statistical experimental designs

1. Pharmaceutical industry: Tablet formulation and chromatographic methods

2. Environmental biotechnology: waste water treatment and bioremediation

3. Chemical industry: optimize reaction conditions for maximum yield

4. Bioprocess development: Optimization of fermentation yield includingenzymes, antibiotics, biopolymers as will as many secondary metabolites

5. Molecular biology: PCR optimization and micro array

What is the next level in experimental design???What is the next level in experimental design???

Evolutionary computationEvolutionary computationNon-model based optimization method, with the ability to search A system with a large number of parameters in directed wayHibbert 1993

Has been applied on formate dehydrogenase enzyme productionWeuster-Botz and Wandery 1995

Has been applied on thermostable Lipase production model with higherDegree of fitness to experimentalData.Abdel-Fattah and Sheta 2003

Genetic Engineering and Biotechnology Research Institute

GEBRIGEBRI