Chapter 7 Prepare the Comparative C
T
Reactions
Prepare the Template
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C
T
Experiments
140
Notes
• Total RNA isolated from liver, kidney, brain, and universal tissues
• One of Applied Biosystems High-Capacity cDNA Reverse Transcription Kits:
Note: The High-Capacity cDNA Reverse Transcription Kit was formerly called the
High-Capacity cDNA Archive Kit.
Prepare the
Template
Use the High-Capacity cDNA Reverse Transcription Kit to reverse-transcribe cDNA
from the total RNA samples. Follow the procedures in the Applied Biosystems High-
Capacity cDNA Reverse Transcription Kits Protocol to:
1. Prepare the RT master mix.
CHEMICAL HAZARD. 10✕ RT Buffer may cause eye, skin,
and respiratory tract irritation. Read the MSDS, and follow the handling
instructions. Wear appropriate protective eyewear, clothing, and gloves.
2. Prepare the cDNA reactions.
3. Perform reverse transcription on a thermal cycler.
PCR Starter
Pack
RT-PCR Grade Water (nuclease-free) AM9935
DNAZap
™
AM9890
Choice of RNA sample preparation products, as
listed above under the qRT-PCR Starter Pack.
See above
High Capacity
cDNA Kit
RNAlater
®
Tissue Collection: RNA Stabilization
Solution
AM7020
RNaseZap
®
Wipes AM9786
Choice of RNA sample preparation products, as
listed above under the qRT-PCR Starter Pack.
See above
miRNA Starter
Pack
Pre-miR
™
miRNA Starter Kit AM1540
Kit Part Number
High-Capacity cDNA Reverse Transcription Kit (200 reactions) 4368814
High-Capacity cDNA Reverse Transcription Kit (1000 reactions) 4368813
High-Capacity cDNA Reverse Transcription Kit with RNase Inhibitor
(200 reactions)
4374966
High-Capacity cDNA Reverse Transcription Kit with RNase Inhibitor
(1000 reactions)
4374967
Kit Contents
Catalog
Number
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