Applied Biosystems 7500 Fast Getting Started

Chapter 3 Prepare the Reactions
Prepare the Standard Dilution Series
47
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Standard Curve
Experiments
Notes
Prepare the Standard Dilution Series
Prepare the standard dilution series using the volumes that were calculated by the 7500
software (“Complete the Reaction Mix Calculations Tab (see page 33)” on page 32).
About the
Example
Experiment
For the standard curve example experiment:
• The standard concentration (stock) is 20,000 copies/µL.
• The volumes calculated in the software are:
Required
Materials
• Water (to dilute the standards)
• Microcentrifuge tubes
• Pipettors
• Pipette tips
• Standard stock
•Vortexer
• Centrifuge
Prepare the
Standard Dilution
Series for the
RNase P Assay
1. Label a separate microcentrifuge tube for each standard:
• RNase P Std. 1
• RNase P Std. 2
• RNase P Std. 3
• RNase P Std. 4
• RNase P Std. 5
Dilution
Point
Source
Source Volume
(µL)
Diluent Volume
(µL)
Total Volume
(µL)
Standard Concentration
(ng/µL)
1 (10,000) Stock 3.63 14.52 18.15 4000.0
2 (5,000) Dilution 1 9.08 9.08 18.15 2000.0
3 (2,500) Dilution 2 9.08 9.08 18.15 1000.0
4 (1,250) Dilution 3 9.08 9.08 18.15 500.0
5 (625) Dilution 4 9.08 9.08 18.15 250.0

Applied Biosystems 7500 Fast Specifications

General

Brand	Applied Biosystems
Model	7500 Fast
Category	Laboratory Equipment
Language	English