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Note: The procedure for this TargetAmp Kit has changed. Please download the latest version of the kit protocol before proceeding.
The TargetAmp™ 2-Round Aminoallyl-aRNA Amplification Kit 1.0*
generates at least 5-million-fold amplification of the poly(A) RNA contained
in a total cellular RNA sample. This amplification is sufficient to generate
microgram amounts of aminoallyl-labeled antisense RNA (AA-aRNA; also
called AA-cRNA) from the total RNA contained in a single cell. The kit
uses an Eberwine-type linear RNA amplification method,1 but
incorporates a number of improvements that significantly increase the
yields of AA-aRNA obtained, while reducing the level of nonspecific amplification
products. The kit is recommended for amplifying mRNA in total RNA from
1 to ~50 cells.
The kit provides all reagents (except reverse transcriptases and RNA
purification columns) for generating AA-aRNA. The entire procedure takes
only ~4 hours of hands-on time and can be completed in 2 days. Multiple
samples can be amplified simultaneously. Once produced, the aminoallyl-aRNA
can be readily labeled using biotin-NHS or fluorescent dye-NHS compounds.
Biotin-NHS is available from Epicentre.
Table 1. The TargetAmp™ Kit 1.0 produces microgram amounts
of AA-aRNA from as little as 10 pg (one cell) of total RNA. Results
represent the average of multiple experiments.
a. Approximately 5% of rat brain total RNA is poly(A) RNA.
b. Approximately 2% of HeLa total RNA is poly(A) RNA.
c. Approximately 1.5%-2% of rat kidney total RNA is poly(A) RNA.
*TargetAmp™ aRNA Amplification Products are covered by intellectual
property licensed to Epicentre Technologies Corporation from Johnson & Johnson
Pharmaceutical Research & Development, L.L.C., and by intellectual property
sublicensed to Epicentre Technologies Corporation from Incyte Corporation.