when am doing ARMS PCR both WT and MT amplified equally? how to minimize WT amplification?
Paul Rutland
you can run temperature gradients of annealing temperatures for each primer set on both a normal and separately a mutant sample.there should be a temperature at which one primer works and the other primer does not work for each primer set and each sample .So if normal primer works at 60c but the mutant primer on a normal sample stops working at 57 c then on other samples a pcr annealing at 59c with both primer sets will only amplify the normal allele. If gradient does not differentiate then redesign the primers or consider dcaps method to create a restriction site to distinguish the alleles
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