Bio-Rad iQ™5 Optical System Software, Version 2.1 - Manual (Page 33)

Section 4 Workshop Module
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9. Click a fluorophore.
10. Click or drag across the plate to define wells with the selected fluorophore and sample
type.
11. Continue defining the remaining wells that will contain the first fluorophore by
changing to any other sample type icons required.
To calculate standard concentrations automatically, click Dilution Series and enter the
upper or lower concentrations and units of the standards, set the dilution factor, and
click Apply Dilution Series.
12. Repeat steps 7–11 for any additional dye layers/fluorophores as required. Remember
that if the Whole Plate Loading box is checked, changes made in standard
concentrations will be applied to all dye layers for that well and extended to all
replicates in the group.
NOTE: To delete a previously defined well, click the Delete All icon, and then click the
well. To delete the selected fluorophore from a previously defined well, click the Delete
Fluorophore icon, and then click the well.
13. Click Save & Exit Plate Editing. The extension .pts will be added automatically
NOTE: Selecting or creating plate setup files that contain fluorophores not compatible
with the connected instrument will result in a warning message displayed at the
bottom of the screen (Figure 4.5). This message indicates that while the plate setup
can be viewed and edited, the plate setup contains fluorophores that cannot be used to
collect new data using this plate setup.
Fig. 4.5. Invalid Plate Setup Error.
4.2.4 Fluorophore Selection
When using the MyiQ2, MyiQ, and iQ5 systems, you may specify as many as five different
fluorophores on a single plate provided they are read through the filters available on the
respective systems. Refer to section 9.2.1 for information about system filter specifications and
recommended fluorophores.
With the iQ5 software, as many as five different sets of wells can be read with the same filter
pair (that is, in the same fluorophore) as shown in Figure 4.6. For example, you may have five
different experiments on the same plate, all using SYBR
® Green, and each experiment will be
treated independently.
NOTE: The software cannot distinguish between fluorophores read by the same filter pair in the
same well; therefore, in all cases, such dyes have to be in separate wells. For example, you may
not put FAM and SYBR
® Green in the same well.