Applied Biosystems StepOne User Manual

DRAFT September 25, 2007 1:07 am, 4376782_Standalone_Installation.fm
Chapter 4 Install the Standalone Layout
Perform the RNase P Experiment
Applied Biosystems StepOne
™
and StepOnePlus
™
Real-Time PCR Systems
Installation, Networking, and Maintenance Guide
60
Notes
Perform the RNase P Experiment
After you install the instrument, confirm the function of the system by running a
TaqMan
®
RNase P Fast Instrument Verification Plate.
Materials
Required
•TaqMan
®
RNase P Fast Instrument Verification Plate
• Powder-free gloves
• Safety glasses
• Centrifuge with plate adapter
When to Perform
the RNase P
Experiment
Applied Biosystems recommends performing an RNase P experiment:
• After installing the system.
• After moving the instrument to another location.
• As needed to confirm the function of the instrument, depending on your laboratory
and regulatory requirements.
Purpose of the
Experiment
The TaqMan
®
RNase P Fast Instrument Verification Plate experiment verifies the
performance of the instrument. The RNase P plate is preloaded with the reagents necessary
for the detection and quantitation of genomic copies of the human RNase P gene (a single-
copy gene encoding the RNase moiety of the RNase P enzyme).
Each well contains:
•1✕ TaqMan
®
Fast Universal PCR Master Mix, No AmpErase
®
UNG
•RNaseP primers
•FAM
™
dye-labeled probe
• Known concentration of human genomic DNA template
The figure below illustrates the arrangement of the standard and unknown populations on
the RNase P plate. The RNase P plate contains five replicate groups of standards (1250,
2500, 5000, 10,000, and 20,000 copies), two unknown populations (5000 and 10,000
copies), and negative control wells.
5K-copy Unknown
Population 1
10K-copy Unknown
Population 2
NC
STD 1250 STD 2500
STD 5000 STD 10000 STD 20000
NC
STD 1.25K
STD 20K
STD 2.5KSTD 5K
STD 10K
10K-copy
Unknown
Population 2
10K-copy
Unknown
Population 2
5K-copy
Unknown
Population 1
5K-copy
Unknown
Population 1
TaqM an
®
48-well RNase P Plate for
the StepOne
™
System
Taq Man
®
96-well RNase P Plate for
the StepOnePlus
™
System

Applied Biosystems StepOne Specifications

General

Brand	Applied Biosystems
Model	StepOne
Category	Laboratory Equipment
Language	English