I know there is a technology called Locked Nucleic Acids, that allows to design shorter PCR oligonucleotides with higher Tm. But how exactly is the Tm calculated, if LNA is incorporated ?
At the moment I can only provide a "rule of the thumb" answer:
"The β-D-LNA modification results in significant increases in melting temperature of up to several degrees per LNA residue."
From: Arnold GrÜnweiler, Roland K Hartmann (2007). Locked nucleic acid oligonucleotides. BioDrugs 21, 235-243.
Matej Lexa yes, that was my impression from other articles too. What confuses me is, that this online calculator adds only 2°C Tm increase per each LNA-modified base. That seems too little to me, and, in fact, is not enough for what I hoped. https://www.genscript.com/tools/oligo-primer-calculation
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