3 quantity standards, Quantity standards -24 – Bio-Rad Quantity One 1-D Analysis Software User Manual

Page 167

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Quantity One User Guide

6-24

Each bar of the histogram is labeled on the X axis with the lane number, and (where
space permits) the band number. The Y axis is labeled with quantitative values. If the
bands in the match group span the entire image window, the histogram will not
include bars for the bands in the left-most lanes because of the space required for
labeling the axis.

To avoid this problem, decrease the size of the image using Zoom Out until there is
blank space between the left side of the window and the first band of the match group.
Redisplay the histogram. If one or more bars are still not included, continue
decreasing the magnification until you can see all the data.

6.3 Quantity Standards

From bands of known quantity, you can generate a calibration curve for determining
the quantities of all the bands in lanes or cells in lane-based arrays. (To quantitate
bands outside of lanes, see Chapter 7.)

Using the Quantity Standards dialog, you plot the quantities of the known bands
against their intensities to generate a calibration curve. You then apply this curve to
unknown bands in the current gel as well as other gels.

To create a calibration curve, the quantities of at least two bands must be known. The
greater the number of known bands and the wider the range of their values, the more
accurate the calibration curve will be.

Note:

The band intensities calculated by Gaussian fitting (see section 5.7, Gauss-
Modeling Bands
) cannot be used in conjunction with Quantity Standards.
However, you can continue to use the trace intensity (i.e., the area under a band’s
intensity profile) to calculate Quantity Standards after you have Gauss-
modeled your bands.

6.3.a Creating and Applying a Set of Quantity Standards

Select Quantity Standards from the Analysis menu. A pop-up box will prompt you
to create a new curve or load a saved calibration curve.

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