Through reading the literature, I got a large number of primers for toxin and antitoxin research. But I want to carry out multiple PCR. How can I determine whether it is necessary to redesign multiple PCR primers? What is its standard? Thank you very much for your answer.
Primers are cheap. Order a few pairs from the literature and test them until you find one that works well.
I think that the number of interactions increases hugely as the number of primers increases in multiplex pcr so you can either spend a lot of time analysinf primer dimer interactions at some site like IDTDNA or just try mixes of primers and troubleshoot those mixes which produce weak bands and primer dimer. If you are collecting random primer sets then it is likely that some will be from old papers and will have low annealing temperatures so it will be worth making these longer at the 5' end to raise the annealing temperature so minimising PD. Also use a hot start polymerase to minimise all primer-primer interactions in your multiplexes
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