PCR Annealing Temperature Estimate

Calculate optimal PCR annealing temperature from primer Tm values and GC content for standard, touchdown, and gradient PCR protocols.

 
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Physical Constants Reference
ConstantSymbolValue
Speed of lightc2.99792458×10⁸
Planck's constanth6.62607015×10⁻³‴
Boltzmann constant1.380649×10⁻²³
Avogadro's numberNₐ6.02214076×10²³
Gravitational constantG6.6743×10⁻¹¹
Gas constantR8.31446
Elementary chargee1.602176634×10⁻¹⁹
Electron massmₑ9.1093837015×10⁻³¹
Proton massmₚ1.67262192369×10⁻²⁷
Fine-structure constantα7.2973525693×10⁻³
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How It Works

Calculate optimal PCR annealing temperature from primer Tm values and GC content for standard, touchdown, and gradient PCR protocols

Each component has a specific meaning:

  • GC content — The gc content recorded for the patient or scenario being assessed.

Note: Interpret the pcr annealing result against the clinical thresholds and context described above.

How to Use

Enter the GC content for the patient or scenario you are assessing. Calculate optimal PCR annealing temperature from primer Tm values and GC content for standard, touchdown, and gradient PCR protocols. Use the pcr annealing result to inform your clinical assessment.

Frequently Asked Questions