METHOD FOR PREPARATION OF DNA LADDER USING PCR AND ITS OPTIMIZATION BY NUMERICAL MODELING THEREOF

摘要

The patent pending describes an easy method for preparing DNA ladder using polymerase chain reaction and, consequently, applying numerical modeling for optimization of PCR yield. In this method (11) oligonucleotide primers have been design to amplify (10) DNA fragments from plasmid DNA with known lengths. Based on the required fragment lengths, three PCR programs have been implemented to give the best amplification results. In order to optimize the PCR yield, numerical modeling methodology has been applied by studying (6) significant variables simultaneously. The optimized PCR yield reached (5) times the basal conditions. In addition, a mathematical equation has been described to correlate the relationship between variables and the PCR yield which saves time and fine chemicals consumption.