The multiplex PCR experiment is to amplify multiple regions of a DNA sequence at the same time by using different primer pairs. Designing feasible primer pairs for multiplex PCR is a tedious task since there are too many constraints to be satisfied. In this paper, a new method for multiplex PCR primer design strategy using genetic algorithm is proposed. The proposed algorithm is able to find a set of suitable primer pairs more efficient and uses a MAP model to speed up the examination of the specificity constraint that is important for gene family sequences. The dry-dock experiment shows that the proposed algorithm finds several sets of primer pairs of gene family sequences for multiplex PCR that not only obey the design properties, but also have specificity.
This study presents a novel application, MultiPrimer, to assist users to design multiplex PCR primers. MultiPrimer adopts an algorithm based on a genetic algorithm and can efficiently solve the multi-objective optimisation problem. A match pattern model is proposed to speed up examination of the specificity constraint. MultiPrimer is able to find a set of primer pairs simultaneously for a set of gene family sequences and multiple targets for a single cDNA sequence. A public version of the MultiPrimer software is available from http://edith.cse.nsysu.edu.tw/new/software/MultiPrimer.htm.
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