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18
June 23, 2003 9:39 am, MFC_QSG_text.fm
To prepare the sample-specific PCR mix:
1. Label a 1.5-mL microcentrifuge tube.
2. Remove a cDNA sample from the freezer. Thaw the sample
by rolling it between your fingers.
3. Gently vortex the sample, then centrifuge the tube.
4. Add the following components to the labeled 1.5-mL
microcentrifuge tube:
Component
cDNA sample
RNase/DNase-free water
®
TaqMan
Universal PCR Master
Mix (2X)
Total
a. Total RNA converted to cDNA. The cDNA sample volume
can vary (up to 50 µL) to achieve a specific sample amount
(in nanograms). Water is added to achieve a total volume
of 50 µL for sample and water.
5. Cap the microcentrifuge tube and thoroughly mix the
solution by gentle vortexing.
6. Centrifuge the tube to eliminate air bubbles from the
mixture.
7. Continue with Loading the Micro Fluidic Cards as described
in Chapter 4 of the user guide for the 7900HT system.
Refer to the guidelines for loading cards and operating the
centrifuge below.
Applied Biosystems Micro Fluidic Card Getting Started Guide
DRAFT
a
(from step 2)
Volume Per Fill
Reservoir (µL)
5
45
50
100
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