Chapter 7 Prepare the Comparative C
T
Reactions
Prepare the Template
141
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C
T
Experiments
Notes
Preparation
Guidelines
When you prepare your own comparative C
T
(∆∆C
T
) experiment Applied Biosystems
recommends:
• That you first extract DNA or RNA from the tissue or sample.
• The following templates:
–
Complementary DNA (cDNA)
– cDNA reverse-transcribed from total RNA
samples using a High-Capacity cDNA Reverse Transcription Kit.
–
RNA
– Purified total RNA or mRNA extracted from tissue or sample.
–
Genomic DNA (gDNA)
– Purified gDNA already extracted from tissue or sample.
For More
Information
For more information on:
• Preparing cDNA templates – Refer to the Applied Biosystems High-Capacity cDNA
Reverse Transcription Kits Protocol. The protocol is not shipped with the High-
Capacity cDNA Reverse Transcription Kits. You can download the protocol from:
http://docs.appliedbiosystems.com/search.taf
• Preparing RNA or gDNA templates – Refer to the protocol for the purification
reagents that you select. To locate Applied Biosystems purification reagents, visit:
http://www.appliedbiosystems.com/
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