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

Author:

Afzal Farwa,Zia Saeeda,Saif RashidORCID

Abstract

AbstractThe 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 56°C for 10 hours and proteinase K enzyme concentration of 10µL. Utilizing these parameters, a yield of 300µL was obtained. Furthermore, other DNA extraction parameters may also be added for a more robust design for obtaining the maximum DNA yield.

Publisher

Cold Spring Harbor Laboratory

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