Applied Biosystems 7500 Fast Getting Started

Chapter 3 Prepare the Reactions
Prepare the Reaction Mix
49
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Standard Curve
Experiments
Notes
Prepare the Reaction Mix
Prepare the reaction mix using the components and volumes that were calculated by the
7500 software (“Complete the Reaction Mix Calculations Tab (see page 33)” on page 32).
Note: The software calculates the volumes for all components for the PCR reactions.
However, when you prepare the reaction mix in this section, include only the master mix,
assay mix, and water. Add the sample or standard when you prepare the reaction plate
(see “Prepare the Reaction Plate” on page 51).
About the
Example
Experiment
For the standard curve example experiment:
• The reaction mix components are:
–TaqMan
®
Universal PCR Master Mix (2✕) or TaqMan
®
2✕ Universal PCR
Master Mix, No AmpErase
®
UNG
– RNase P Assay Mix (20✕)
– Water
• The volumes calculated in the software are:
Note: The sample or standard is not added at this time.
Required
Materials
• Microcentrifuge tubes
• Pipettors
• Pipette tips
• Reaction mix components (listed above)
• Centrifuge
Prepare the
Reaction Mix
CHEMICAL HAZARD . TaqMan
®
Universal PCR Master Mix (2 ✕ )
may cause eye and skin irritation. Exposure may cause discomfort if swallowed or inhaled.
Read the MSDS, and follow the handling instructions. Wear appropriate protective eyewear,
clothing, and gloves.
Component Volume for 1 Reaction (µL)
Master Mix (2.0✕) 12.50
Assay Mix (20.0✕)1.25
H
2
O8.75
Total Volume 22.50

Applied Biosystems 7500 Fast Specifications

General

Brand	Applied Biosystems
Model	7500 Fast
Category	Laboratory Equipment
Language	English