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Invalid characters detected. Only A, T, G, C allowed.
Invalid characters detected. Only A, T, G, C allowed.
Primer Advisory:
Thermodynamic Parameters:
ΔH: kcal/mol
ΔS (salt-corrected): cal/K·mol
Predicted Tm: °C
Stacking interactions dominate stability. GC pairs contribute both stronger hydrogen bonding and stacking.
Fraction melted: 0.0%
Absorbance (260nm): 1.00
Tm: --.- °C
Effective Efficiency: 0.00
Final Yield: 0 copies
WT + Probe Tm: --
Mut + Probe Tm: --
ΔTm: --
Specificity: --
RNA-DNA Tm: --
Mismatches: 0
High mismatch penalty in seed region!
Energy: 0 kcal/mol
Base Pairs: 0
Adenine (A)
Thymine (T)
Guanine (G)
Cytosine (C)
DNA Thermodynamics
Salt: Positive ions (Na+) shield the negative charges on the DNA phosphate backbone, reducing repulsion and stabilizing the double helix (increases Tm).
GC Content: G-C pairs have 3 hydrogen bonds and stronger base-stacking interactions than A-T pairs, requiring more energy to break (increases Tm).
Length: Longer sequences have more cooperative interactions. Short primers melt over a broader temperature range with less sharp transitions.