Not a Constant Value: Understanding Tm
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In the latest issue of DECODED, the quarterly newsletter form Integrated DNA Technologies, resident expert on nucleic acid thermodynamics, Dr Richard Owczarzy has introduced researchers to a new approach for calculating nucleic acid melting temperature (Tm).
Exploring the impact of environmental conditions on Tm, the article explains how researchers can use these calculations to enhance the performance of their hybridization-based protocols.
Oligonucleotide hybridization forms the basis of many molecular biology techniques, from PCR and NGS through to in situ hybridization and gene knockdown studies, and knowing the exact Tm is vital when designing these experiments.
Dr Owczarzy explains how many researchers simplify their calculations of Tm as a fixed characteristic based on sequence composition.
However, Tm is also dependent on experimental conditions, including oligo concentration, the ionic environment and Mg2+ availability, as well as sequence mismatches and SNPs.
IDT has therefore developed a free online Tm calculator, delivering the most accurate calculations of Tm.
