Mathematical modeling using box-behnken response surface design for optimal DNA yield with variable parameters of time, temperature, and proteinase K concentration

Abstract

The current research aims to explore an enzyme-assisted inorganic DNA extraction protocol to obtain maximum yield from whole blood. We considered various operating conditions such as incubation temperature and time along with proteinase K enzyme concentration. To model and simulate the extraction process, Box-Behnken Design (BBD) was employed with three factors and its five levels, coupled with the desired functional response surface methodology (RSM). RSM entails fitting a mathematical model for experimental data to optimize a response variable with Box-Behnken Design. The best simulation settings yielded the maximum DNA at an incubation temperature of 56C for 10 hours and proteinase K enzyme concentration of 10μL. Utilizing these parameters, a yield of 300ng/μL was obtained. Furthermore, other DNA extraction parameters may also be added for a more robust design for obtaining the maximum DNA yield.