Analyze The Experiment - Applied Biosystems StepOne Installation, Networking, And Maintenance

Chapter 4 Install the Standalone Layout
Analyze the RNase P Experiment

Analyze the Experiment

Confirm the
Results of the
Analysis
TaqMan
Notes
68
Review the data to confirm the results of the experiment.
1. In the Analysis screen of the RNase P Run wizard, confirm the status of the run:
• Passed – The instrument passed the RNase P run. Go to
• Failed – The instrument failed the RNase P run. Go to
experiment for outliers.
If the run fails, the automated analysis may have included outliers that caused the
initial analysis to fail. Experimental error may cause some wells to be amplified
insufficiently or not at all. These wells typically produce C
significantly from the average for the associated replicate wells. If included in the
calculations, these outlying data (outliers) can result in erroneous measurements.
2. In the Amplification Plot, select Ct vs. Well from the Plot Type dropdown menu.
3. Confirm the uniformity of each replicate population on the RNase P plate (controls,
standards, and unknowns) by comparing the groupings of C
In the plate layout, select the wells containing the 10,000-copy unknown
a.
population:
™
• StepOne
• StepOnePlus
5K-copy 5K-copy
Unknown Unknown
Population 1 Population 1
10K-copy 10K-copy
Unknown Unknown
Population 2 Population 2
®
48-well RNase P Plate for
™
the StepOne System
In the plot, confirm that the C
b.
The numbers on the X-axis of the plot correspond to the wells of the
Note:
reaction plate. Beginning with well A1, the wells are numbered from left-to-
right, and top-to-bottom.
Applied Biosystems StepOne
System – Wells from columns 1, 2, 6, 7, and 8 in rows D, E, and F
™
System – Wells rows F, G, and H
NC
STD 5000
TaqMan
the StepOnePlus
s of the replicate population are equivalent.
T
™
and StepOnePlus
Installation, Networking, and Maintenance Guide
step 5 on page
step 2 to screen the
values that differ
T
values:
T
5K-copy Unknown
Population 1
STD 1250 STD 2500
STD 10000 STD 20000
10K-copy Unknown
Population 2
®
96-well RNase P Plate for
™
System
™
Real-Time PCR Systems
70.