Applied Biosystems ViiA 7 Real, Краткое руководство пользователя

Страница: 6 / 22

6
Perform an experiment
Applied Biosystems ViiA
™
7 Real-Time PCR System Quick Reference Guide
Design the experiment
1
Define experiment name
and type (Experiment
Properties screen) 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
T
(∆∆C
T
) , Melt
Curve , Genotyping , or Presence/Absence
2
Select the reagents
(Experiment Properties
screen) Select reagents to use in the experiment:
Note: If you select SYBR
®
Green as the reagent, then you have the option of including a melt curve
for that experiment.
3 Define the instrument
run properties
(Experiment Properties
screen) • Ramp speed: Standard or Fast
• (Genotyping and Presence/Absence) Collect data during Pre-PCR Read , Amplification , or Post-
PCR Read
• (Experiments using SYBR
®
Green) Select whether or not to include a melt curve.
• (Melt Curve) Select whether or not to include PCR.
4 Define targets, samples,
biological replicates,
and dye (Define screen) • (All experiments except Genotyping) Define Targets.
• (Genotyping) Define SNPs.
• (All experiments) Define Samples.
• (Optional) (Standard Curve, Relative Standard Curve, and Comparative C
T
) Define Biological
Replicate Groups.
• (All experiments except Presence/Absence) Select the Passive Reference dye.
• (Relative Standard Curve and Comparative C
T
) Select the Reference Sample and the Endogenous
Control.
5
Assign targets and
samples (Assign screen) • (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
T
) Assign Biological
Replicate Groups to wells.
Experiment type Reagent options
Standard Curve TaqMan
®
Reagents , SYBR
®
Green Reagents , or Other
Relative Standard Curve TaqMan
®
Reagents , SYBR
®
Green Reagents , or Other
Comparative C
T
TaqMan
®
Reagents , SYBR
®
Green Reagents , or Other
Melt Curve SYBR
®
Green Reagents or Other
Genotyping TaqMan
®
Reagents or Other
Presence/Absence TaqMan
®
Reagents or Other