Formamide concentration in percentage (fmdmethod="concentration") or molar (fmdmethod="molar").
DMSOfactor
Coefficient of Tm decreases per percent DMSO. Default=0.75 von Ahsen N (2001) <PMID:11673362>. Other published values are 0.5, 0.6 and 0.675.
fmdmethod
"concentration" method for formamide concentration in percentage and "molar" for formamide concentration in molar
fmdfactor
Coefficient of Tm decrease per percent formamide. Default=0.65. Several papers report factors between 0.6 and 0.72.
ptGC
Percentage of GC(%).
Details
fmdmethod = "concentration"
Correction = - factor*percentage_of_formamide
fmdmethod = "molar"
Correction = (0.453*GC/100 - 2.88) x formamide
References
von Ahsen N, Wittwer CT, Schutz E , et al. Oligonucleotide melting temperatures under PCR conditions: deoxynucleotide Triphosphate and Dimethyl sulfoxide concentrations with comparison to alternative empirical formulas. Clin Chem 2001, 47:1956-C1961.