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Applied Biosystems StepOne User Manual

Applied Biosystems StepOne
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DRAFT
September 25, 2007 1:07 am, 4376782_Maintain_the_System.fm
Chapter 6 Maintain the System
Regular Maintenance
Applied Biosystems StepOne
and StepOnePlus
Real-Time PCR Systems
Installation, Networking, and Maintenance Guide
94
Notes
Perform a Background Calibration
Perform a background calibration monthly, or as often as necessary, depending on
instrument use.
Materials
Required
Background plate from the spectral calibration kit for your instrument
:
StepOne
Real-Time PCR System Spectral Calibration Kit (PN 4371433)
StepOnePlus
Real-Time PCR System Spectral Calibration Kit (PN 4371435)
Note: Alternatively, create a background plate as described in “How to Create a
Background Plate” on page 99.
Powder-free gloves
Safety glasses
Centrifuge with reaction plate adapter
StepOne system cable, yellow (from the system packing kit)
Calibration
Guidelines
Make sure the centrifuge you use is clean. Before centrifuging, wipe down the
bucket using a tissue.
Handle the calibration plates with care to prevent contamination. To avoid
contaminating the reaction plate, do not place plates on a lab bench. Always return
the calibration plate to its original bag.
Purpose of the
Calibration
A background calibration measures the level of background fluorescence in the
instrument. During a background calibration run, the system:
Performs continuous reads of a background plate containing PCR buffer for 10 min
at 60 ° C.
Averages the spectra recorded during the run and extracts the resulting spectral
component to a calibration file.
The StepOne
software uses the calibration file during subsequent runs to remove the
background fluorescence from the run data.
Background Fluorescence
Fluorescence data collected by the system includes a fluorescent spectral component
inherent to the system, commonly referred to as background fluorescence. This
background fluorescence is a composite signal found in all signal data that consists of
fluorescence from several sources, including:
Background electronic signal
Contaminants in the sample block(s)
The plastic consumable (plates and caps)

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Applied Biosystems StepOne Specifications

General IconGeneral
BrandApplied Biosystems
ModelStepOne
CategoryLaboratory Equipment
LanguageEnglish

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