Design The Experiment - Applied Biosystems ViiA 7 Real Quick Reference Manual

Perform an experiment

Design the experiment

1
Define experiment name
and type (Experiment
Properties screen)
2
Select the reagents
(Experiment Properties
screen)
3 Define the instrument
run properties
(Experiment Properties
screen)
4
Define targets, samples,
biological replicates,
and dye (Define screen)
5
Assign targets and
samples (Assign screen)
6
Define the following properties:
• Experiment name
• User name of the experiment owner
• (Optional) Barcode for the plate or array card
• (Optional) Comments
• Block type: 384-well, Array Card, 96-well (0.2mL), or Fast 96-well (0.1mL)
• Experiment type: Standard Curve, Relative Standard Curve, Comparative C
Curve, Genotyping, or Presence/Absence
Select reagents to use in the experiment:
Experiment type
Standard Curve
Relative Standard Curve
Comparative C
T
Melt Curve
Genotyping
Presence/Absence
®
Note:
If you select SYBR Green as the reagent, then you have the option of including a melt curve
for that experiment.
• Ramp speed: Standard or Fast
• (Genotyping and Presence/Absence) Collect data during Pre-PCR Read, Amplification, or Post-
PCR Read
• (Experiments using SYBR
• (Melt Curve) Select whether or not to include PCR.
• (All experiments except Genotyping) Define Targets.
• (Genotyping) Define SNPs.
• (All experiments) Define Samples.
• (Optional) (Standard Curve, Relative Standard Curve, and Comparative C
Replicate Groups.
• (All experiments except Presence/Absence) Select the Passive Reference dye.
• (Relative Standard Curve and Comparative C
Control.
• (All experiments except Genotyping) Assign targets, tasks, and samples to wells. Tasks include
Unknown, Standard, Positive, and Negative controls, depending on experiment type.
• (Genotyping) Assign SNP assays, tasks, and samples to wells.
• (Standard Curve and Relative Standard Curve) Define and set up standards: Click Define and Set
Up Standards, select a target, define the standard curve, then select and arrange wells for the
standards.
• (Optional) (Standard Curve, Relative Standard Curve, and Comparative C
Replicate Groups to wells.
Applied Biosystems ViiA
Reagent options
®
TaqMan Reagents, SYBR
®
TaqMan Reagents, SYBR
®
TaqMan Reagents, SYBR
®
SYBR Green Reagents or Other
®
TaqMan Reagents or Other
®
TaqMan Reagents or Other
®
Green) Select whether or not to include a melt curve.
) Select the Reference Sample and the Endogenous
T
™
7 Real-Time PCR System Quick Reference Guide
(∆∆C
T
®
Green Reagents, or Other
®
Green Reagents, or Other
®
Green Reagents, or Other
) Define Biological
T
) Assign Biological
T
), Melt
T