Applied Biosystems 7500 - Page 70

Chapter 3 Prepare the Reactions
Prepare the Sample Dilutions
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Standard Curve
Experiments
46
Notes
3. Add the required volume of sample stock to each tube:
4. Vortex each diluted sample for 3 to 5 seconds, then centrifuge the tubes briefly.
5. Place the diluted samples on ice until you prepare the reaction plate.
Preparation
Guidelines
When you prepare your own standard curve experiment:
• Sample dilutions may be necessary because the sample volume is limited to 10% of
the total reaction volume in the 7500 software. You must perform the sample
dilutions before adding the samples to the final reaction mix.
• For optimal performance of TaqMan
®
Gene Expression Assays or Custom TaqMan
®
Gene Expression Assays, use 10 to 100 ng of cDNA template per
20-µL reaction. For Fast reagents, Applied Biosystems recommends 10 ng.
• Use TE buffer or water to dilute the sample.
For More
Information
For more information on Applied Biosystems assays, refer to the:
• TaqMan
®
Gene Expression Assays Protocol
• Custom TaqMan
®
Gene Expression Assays Protocol
Tube Sample Name Sample Volume (µL)
1 Population 1 1.0
2 Population 2 1.0