Perform The Rnase P Experiment - Applied Biosystems StepOne Installation, Networking, And Maintenance

Chapter 3 Install the Colocated Layout

Perform the RNase P Experiment

Perform the RNase P Experiment

Materials
Required
When to Perform
the RNase P
Experiment
Purpose of the
Experiment
Notes
38
After you establish system communication, confirm the function of the system by
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running a TaqMan RNase P Fast Instrument Verification Plate.
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• TaqMan RNase P Fast Instrument Verification Plate
• Powder-free gloves
• Safety glasses
• Centrifuge with reaction plate adapter
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.
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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:
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• 1✕ TaqMan Fast Universal PCR Master Mix, No AmpErase
• RNase P primers
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• 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 (NC).
5K-copy 5K-copy
Unknown Unknown
Population 1 Population 1
10K-copy 10K-copy
Unknown Unknown
Population 2 Population 2
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TaqMan 48-well RNase P Plate for
™
the StepOne System
Applied Biosystems StepOne
®
UNG
5K-copy Unknown
Population 1
NC STD 1250
STD 5000 STD 10000
10K-copy Unknown
Population 2
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TaqMan 96-well RNase P Plate for
™
the StepOnePlus System
™ ™
and StepOnePlus Real-Time PCR Systems
Installation, Networking, and Maintenance Guide
STD 2500
STD 20000