Techne PrimeQ - Manual (Page 122)

122
Calculating the quantification cycle
Fit points are a defined number of fluorescent points found on the curve just above the crossing
line. The points are used in a linear regression calculation and the interception of the linear
regression through the fit points and crossing cursor provides a value for the quantification cycle.
This will be a fractional number calculated for each well. All the user has to do is set the number of
fit points to be included in the calculation and the software does the rest.
Overview of procedure:
3.14.2.5 First derivative maximum
Overview of procedure:
Procedure
• In the Cq calculation window of the Quantification Wizard, click First Derivative Maximum
and then Next.
As this method calculates Cq by defining the point of fastest rate of change in fluorescence, no
noise threshold or crossing cursor has to be set. Therefore, the wizard links straight through to the
Report Options.
3.14.2.6 Report Options and Summary
• Clicking Next leads through to the Report Options screen. Click the appropriate boxes to
select which data is to be displayed in the report.
• Click Next to view a summary of the setup.
• Click Back to change any settings or Cancel to abort the procedure.
• Click Finish to complete the setup.
Fluorescent data for number of fit points above crossing line
1. Software calculates a best-fit log-linear fluorescence curve
through selected points using linear regression (y = mx + c)
2. Intersection of this line through the crossing line calculated as a
fractional cycle number. This is the crossing point (Cp)
Cp values for each well
Fluorescent data for number of fit points above crossing line
1. Software calculates a best-fit log-linear fluorescence curve
through selected points using linear regression (y = mx + c)
2. Intersection of this line through the crossing line calculated as a
fractional cycle number. This is the Cq.
Cq values for each well
Fluorescent data for number of fit points above crossing line
1. Software calculates a best-fit log-linear fluorescence curve
through selected points using linear regression (y = mx + c)
2. Intersection of this line through the crossing line calculated as a
fractional cycle number. This is the crossing point (Cp)
Cp values for each well
Fluorescent data for number of fit points above crossing line
1. Software calculates a best-fit log-linear fluorescence curve
through selected points using linear regression (y = mx + c)
2. Intersection of this line through the crossing line calculated as a
fractional cycle number. This is the Cq.
Cq values for each well