User`s manual

Alpha Innotech User’s Manual, version 4
5-72
Calibration Curves for Quantitative PCR:
The button in the Spot Denso toolbox labeled STD CURVE opens a set of tools that
create a calibration curve for applications such as quantitative PCR.
The calibration curve functions allow quantitation of the bands on a gel based on a set of
standards. A minimum of two standard bands must be input, but the accuracy of the
calibration curve increases as the number of standard bands and their range of values
increase.
Defining the Bands as Objects
Before generating a standard curve, define the bands using the object drawing tools in the
Spot Denso toolbox. As described in Drawing Objects above, the size of an object can
influence the integrated density value. Therefore, we recommend using the same sized
box, ellipse or freehand drawing when defining a standard curve. (See Copying Objects
above for instructions.)
Specifying the Quantitation Units
Once the objects are defined, click on the STD CURVE button. The Standard Curve
Toolbox will appear. Enter the units in which the results should be reported (e.g. ng, ul,
pg, %).
The Standard Curve Toolbox
Press the <Enter> key on the keyboard after typing the units. This step is very
important. If the Enter key is not hit, an error message will appear:
Designating the Standard Bands