Applied Biosystems StepOne - Background Calibration Failure

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
110
Notes
How to Calibrate
a Custom Dye
Materials Required
• Centrifuge with reaction plate adaptor
• Custom dye(s)
• Deionized water
• MicroAmp Optical Adhesive Film or MicroAmp
™
8-Cap Strip (Flat Caps only)
•MicroAmp
™
Optical Reaction Plate of the size appropriate for your instrument
• Pipettors and pipette tips (200-µL and 1000-µL)
• Powder-free gloves
• Safety glasses
• Tubes (2-ml and 10-ml)
Prepare the Custom Spectral Calibration Plate
1. Create a calibration plate for the custom dye:
a. Prepare 1.5 mL of the custom dye at the desired concentration.
b. Pipette 30 µL of the diluted custom dye to all wells of a reaction plate.
c. Seal the wells of the reaction plate using an adhesive film or optical flat caps.
d. Vortex the calibration plate for 5 sec.
e. Centrifuge calibration plate for 2 min at less than1500 rpm.
IMPORTANT! The calibration plate must be well mixed and centrifuged.