Calcolatore Tm Primer PCR
Calcola la temperatura di melting dei primer per ottimizzare le condizioni di PCR.
Y-Intercept (1/Vmax)
0.010000
Y-Intercept (1/Vmax) vs Vmax
Formula
## Lineweaver-Burk Plot The Lineweaver-Burk (double reciprocal) plot linearizes the Michaelis-Menten equation by plotting 1/v vs 1/[S]. ### Formula **1/v = (Km/Vmax) × (1/[S]) + 1/Vmax** This is a straight line with slope = Km/Vmax, y-intercept = 1/Vmax, and x-intercept = -1/Km. While historically important, nonlinear regression is preferred today because Lineweaver-Burk overweights low-concentration data.
Esempio Risolto
An enzyme with Vmax = 100 umol/min and Km = 50 uM.
- 01Y-intercept = 1/100 = 0.01
- 02Slope = 50/100 = 0.5
- 03X-intercept = -1/50 = -0.02
Domande Frequenti
Why is Lineweaver-Burk still taught?
It provides a visual way to distinguish inhibition types: competitive inhibition changes the slope but not the intercept, while noncompetitive changes the intercept but not the slope. These patterns are easily seen on the LB plot.
What are the disadvantages?
The double reciprocal transformation distorts experimental error, giving excessive weight to the least precise measurements (at low [S]). Nonlinear least-squares fitting of the untransformed data is statistically superior.
What other linearization methods exist?
Eadie-Hofstee (v vs v/[S]), Hanes-Woolf ([S]/v vs [S]), and direct linear plots. Each has different error-weighting properties. Eadie-Hofstee is considered the best linear method.
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