How to set up TETRA-ARMS PCR annealing temperatures?

04 August 2021 1 2K Report

Hello everyone .I’m trying to do TETRA ARMS PCR. My outer Forward and Reverse primers have TM: 58 and TM: 50 and inner Forward and Reverse primers have TM: 52 and TM: 48. How can I setup at annealing temperatures?

Can anyone help me?

Paul Rutland

I would redesign the primers so that they all worked at about 60c. Then run gradient pcrs on the primer sets separately on a heterozygous dna sample and see which temperature range gives an amplimer of good intensity for all of the primer sets and use a temperature in the middle of that range

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