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Guidelines for
Use only total RNA samples to generate cDNA for use with the
Micro Fluidic Card. The 18S RNA endogenous control assay cannot
Preparing RNA
+
accurately evaluate cDNA generated from poly A
RNA samples
because most of the rRNA has been removed from them.
See the Applied Biosystems product insert included
IMPORTANT!
with your reagent kit for specific information on the recommended
template for use with the kit.
The High Capacity cDNA Archive Kit Protocol (PN 4322169) also
contains additional guidelines for preparing the RNA template.
Starting Amount
The starting amount of total RNA that you need to convert to cDNA
depends on which Micro Fluidic Card Configuration you intend to
of Total RNA
use. For example, Configuration 1 allows you to load eight different
samples into the eight fill reservoirs on the card, whereas
Configuration 10 requires you to load the same sample into all eight
fill reservoirs on the card. Refer to the assay information file (AIF)
on the Micro Fluidic Card Information CD to confirm the number of
fill reservoirs per sample.
Based on guidelines for Micro Fluidic Cards (discussed in
"Amount
of cDNA in the PCR Mix" on page
17), you should load between
1–100 ng of total RNA converted to cDNA per fill reservoir.
After thermal cycling, store all cDNA samples at -15 to -25 ° C. To
Storing cDNA
minimize repeated freeze-thaw cycles of cDNA, Applied Biosystems
recommends that you store your cDNA samples in aliquots.
14
Applied Biosystems Micro Fluidic Card Getting Started Guide
DRAFT
June 23, 2003 9:39 am, MFC_QSG_text.fm
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