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How do I calculate the Tm of my oligo if I want to incorporate mismatches?
The free, IDT online OligoAnalzyer tool, accessed from the SciTools tab on the IDT website (www.idtdna.com/scitools), allows researchers to set mismatches. Enter your sequence into the input box, and then click the Tm Mismatch button. Click on the base you wish to change and then set the mismatch base using the drop down menu. After clicking <calculate>, the exact match and mismatch Tms will be displayed.