BIO RAD CFX384, Instruction Manual

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Plates
46
Well Loading Controls
A plate file contains information about the contents of each well loaded with sample for an
experiment. After the run, the software links the well contents to the fluorescence data
collected during the protocol and applies the appropriate analysis in the Data Analysis window.
For example, wells loaded with standard sample type are used to generate a standard curve.
When setting up a gene expression experiment, consider the following guidelines for well
contents:
• Target Name. One or more targets of interest (genes or sequences) in each loaded well.
Each target is assigned to one fluorophore
• Sample Name. One identifier or condition that corresponds to the sample in each
loaded well, such as “0 hr”, “1 hr”, or “2 hr”
TIP: Target names and sample names must match between wells to compare data
in the Gene Expression tab in the Data Analysis window. Each name must contain
the same punctuation and spacing. For example, “Actin” is not the same as
“actin”, and “2hr” is not the same as “2 hr”. To facilitate consistency in names,
enter them in the Target and Sample Names Libraries in the Plate tab in the User
Preferences window (page 125).
Select a well to load contents into by left clicking with the mouse pointer in the plate view.
Hold down the mouse button and drag to select multiple wells. The buttons and lists on the
right side of the plate view include all the options needed to load the wells (Table 17).
Table 17. Options for loading the plate and wells in the Plate Editor
Option Function
After selecting wells, the Sample Type must be loaded
first. Select a
Sample Type from the pull-down menu to
load it in the selected wells, including Unknown,
Standard, NTC (no template control), Positive Control,
Negative Control, and NRT (no reverse transcriptase)
Click a Load box to add a fluorophore to the selected
wells; each fluorophore corresponds to a target name. To
add fluorophores to the Load list, select them in the
Select Fluorophores window
For gene expression analysis or to distinguish between
multiple targets, select a name in the
Target Name pull-
down menu and press the Enter key to load the target
name in the well. To delete a target name, select it, press
the
Delete key, and press the Enter key
TIP: To add a new target name to the pull-down menu in
the current plate only, type a name in the pull-down box
and press the
Enter key