Why is the Tm provided on your oligonucleotide specification sheets different from the calculated Tm I obtained when I designed my oligos?

Jul 28, 2013, 14:01 PM

Oligonucleotides designed using our online SciTools® programs (e.g., the PrimerQuest Primer Design ToolRealTime PCR Design Tool or the OligoAnlayzer® Tool) are ordered for a wide variety of applications, and not solely for PCR. The Tm shown on the spec sheets is calculated based on default conditions of 50 mM Na+ without taking into consider­ation Mg2+ and dNTP concentrations, both which influence Tm. To determine the correct Tm for your oligo under your specific reaction conditions, enter the sequence into the on­line OligoAnalyzer® Tool, making sure to change the defaults (32 mM Na+, 3 mM Mg2+, and 0.8 mM dNTPs) to your reactions conditions.

  • SciTools Applications
  • Oligo Characteristics
  • scitools
  • primerquest
  • Tm
  • oligoanalyzer
  • RealTime