Chapter 2 Design the Standard Curve Experiment
Set Up the Samples
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Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Standard Curve
Experiments
Notes
b.
In the Place Negative Controls in drop-down list, select
Upper Left
(default).
10. Click Next.
Design
Guidelines
When you design your own standard curve experiment:
• Identify each sample with a unique name and color. You can enter up to 100
characters in the Sample Name field.
• Enter the number of replicates (identical reactions) to set up. Applied Biosystems
recommends three replicates for each sample reaction.
• Enter the number of negative control reactions to set up. Applied Biosystems
recommends three negative control reactions for each target assay.
• You can set up biological replicate groups (see “Set Up Biological Replicate
Groups”). Biological replicates allow you to assess the representative nature of your
results as they relate to the population being studied. Inclusion of biological
replicates can give insight into any natural variation that is present within the
population.
• Select which targets to test in the samples:
– Select
All Sample/Target Reactions
to test all targets in all samples.
– Select
Specify Sample/Target Reactions
to specify the targets to test in each sample.
IMPORTANT!
When you use the Design Wizard to set up a standard curve
experiment, you can set up only singleplex reactions (amplification and detection of
one target per well). If you want to set up a standard curve experiment with multiplex
reactions (amplification and detection of two or more targets per well), design your
experiment using Advanced Setup (see page 98) instead of the Design Wizard.
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