I´m designing some new primers for SYBR Green Real-time PCR, and I would like to be sure that there is a low probability of secondary structures formation in my primers and also in the template. Does anyone have some suggestions?
Mfold for predicting template and DINAmelt http://mfold.rna.albany.edu/?q=DINAMelt/Two-state-melting for primer structures and complementarity. You could also try designing primers using the IDT website, as it will provide you with structures as well. More information also at http://www.qpcrexpert.com/assay-design.html
Dear all,
Thanks for your answers.
I used IDT Oligo Analyzer, but I cannot find the way to set up the temperature for making the complementarities analysis. Fortunately this can be easily done in Mfold and DINAMelt
Is use VectorNTI and DNAfold http://mfold.rna.albany.edu/?q=mfold/dna-folding-form
may i know why i couldnt access the mfold website? is there any issue on the website?
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