Hello everyone
I've been having trouble getting results for a particular set of primers, but I can't get any bands, so I decided to do a contact PCR, but the result was the same, no visual bands at all.
I used 25 runs with a starting temperature of 67°C to 55°C, reducing 0.5°C per run, and different samples from different strains of the organism my primers target.
I will appreciate your help :)
you may not be running enough cycles. At the higher temperatures the primers are not annealing so you may only have 10 cycles fully working. Try adding another 20 cycles at your lowest annealing temperature and hopefully there will be visible bands
I agree with Dr Rutland, you need to add more cycles to your pcr reaction. And be sure your nucleic acid samples are good.
Joseph Japhet, Paul Rutland Thank you so much both I will try and let you know the results soon :)
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