DRAFT
June 23, 2003 9:39 am, MFC_QSG_text.fm
18 Applied Biosystems Micro Fluidic Card Getting Started Guide
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:
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.
Component
Volume Per Fill
Reservoir (µL)
cDNA sample
a
(from step 2)
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
RNase/DNase-free water 45
Taq Man
®
Universal PCR Master
Mix (2X)
50
Tot al 1 00