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When performing normalization of real time PCR results, I found two ways of doing it:

1. In my lab they follow the next layout: $efficiency^{-(CT\ interest\ gene - CT\ housekeeping)}$ each time controls with controls and treatments with treatments. Then they divide the all values (treated and controls) each with controls.

2. On the other hand I found another way to normalize that follows this steps: $efficiency ^{(control\ CT - treatment\ CT)}$. Then you make the same for the normalizing gene and divide the first by the last.

The values obtained are similar but not exact. Also I noticed that first algorithm gives rather different values for most diluted when performing a standard curve. Which is correct?

When performing normalization of real time PCR results, I found two ways of doing it:

1. In my lab they follow the next layout: $\text{Efficiency}^{-(CT\ _{\large\text{interest gene}} - CT _{\large\text{ housekeeping}})}$ each time controls with controls and treatments with treatments. Then they divide the all values (treated and controls) each with controls.

2. On the other hand I found another way to normalize that follows this steps: $\text{Efficiency}^{(CT_{\large\text{control}} - CT_{\large\text{treated}})}$. Then you make the same for the normalizing gene and divide the first by the last.

The values obtained are similar but not exact. Also I noticed that first algorithm gives rather different values for most diluted when performing a standard curve. Which is correct?

When performing normalization of real time PCR results, I found two ways of doing it:

1. In my lab they follow the next layout: $efficiency^{-(CT\ interest\ gene - CT\ housekeeping)}$ each time controls with controls and treatments with treatments. Then they divide the value of control with the non-treated value.

2. On the other hand I found another way to normalize that follows this steps: $efficiency ^{(control\ CT - treatment\ CT)}$. Then you make the same for the normalizing gene and divide the first by the last.

The values obtained are similar but not exact. Also I noticed that first algorithm gives rather different values for most diluted when performing a standard curve. Which is correct?

When performing normalization of real time PCR results, I found two ways of doing it:

1. In my lab they follow the next layout: $efficiency^{-(CT\ interest\ gene - CT\ housekeeping)}$ each time controls with controls and treatments with treatments. Then they divide the all values (treated and controls) each with controls.

2. On the other hand I found another way to normalize that follows this steps: $efficiency ^{(control\ CT - treatment\ CT)}$. Then you make the same for the normalizing gene and divide the first by the last.

The values obtained are similar but not exact. Also I noticed that first algorithm gives rather different values for most diluted when performing a standard curve. Which is correct?