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Applied Biosystems StepOne User Manual

Applied Biosystems StepOne
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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

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Applied Biosystems StepOne Specifications

General IconGeneral
BrandApplied Biosystems
ModelStepOne
CategoryLaboratory Equipment
LanguageEnglish

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