Large scale gene expression studies
require reliable high-throughput methods to synthesize and then
purify high quality RNA from a large number of samples. Although the
MEGAscript RNAi Kit includes reagents for both dsRNA production and
clean up, it is not optimized for clean up in a 96 well plate
format. Thus for high-throughput applications, we recommend
combining Ambion's MEGAscript High Yield Transcription Kit with
MEGAclear-96, which is designed specifically for high-throughput and
rapid clean up of large scale transcription reactions.
To demonstrate the utility of this
approach, Ambion developed an automation protocol based on the
MEGAclear technology to purify RNA from high yield in
vitro transcription reactions. Two dsRNAs specific for the
U2AF50 gene product, synthesized using MEGAscript, were
purified using the MEGAclear-96 Kit on the Packard
MultiPROBE II HT or the Biomek 2000 platforms. The activities of the
purified dsRNAs were then tested in Drosophila L2 cells. As
shown in Figure 1, a silencing effect similar to dsRNAs prepared and
purified with the MEGAscript RNAi Kit was observed.
The MEGAclear-96 Kit can clean up as
many as 96 samples in just half an hour. This procedure can be integrated
with your favorite robotic platform or used with manual methods for many
downstream applications, including RNA interference, aRNA amplification,
array hybridization and translation studies.
Figure 1. Comparison
of dsRNA Prepared and Purified with the MEGAscript'"/>
Source:
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