Number of standard replicates-For more accurate efficiency measurements, include replicates to decrease the effects of pipetting inaccuracies. Range of standard quantities-For more accurate and precise efficiency measurements, use a broad range of standard quantities, 5 to 6 logs (10 5 to 10 6 fold).

Factors that affect amplification efficiency: A slope that is close to в€’3.3 indicates optimal, 100% PCR amplification efficiency. SlopeВ - The amplification efficiency is calculated using the slope of the regression line in the standard curve. TargetВ - Name of the target sequence that is currently displayed in the plot. Standards-Data points for wells that contain samples with known quantities of the target sequence. Where m is the slope, b is the y-intercept, and Qty is the standard quantity. Regression line-The best-fit line from the standard curve, which is calculated as: Unknowns-Data points for wells that contain samples with unknown quantities of the target sequence. Target dropdown listВ - Selects the data from the target sequence that is displayed by the plot. Allows you to adjust the view options for the plot.

Zooms out the plot to show all data points. Allows you to click and manually move the position of the plot. Select individual data points from the plot. Toolbar-Contains the following tools for controlling the plot:

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