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
vs. the MEGAscript Kit Followed by MEGAclear-96 .
Western blot analysis was performed on Drosophila
L2 cells 72 hr post treatment with 1 or 10 nM of the indicated U2AF50-specific
dsRNA. RNAi Kit: dsRNA prepared and purified using the MEGAscript
RNAi Kit. Mega 96: dsRNA prepared with the MEGAscript Kit (Cat
#1334) and purified with MEGAclear-96 (Cat. #1909).
For prices and availability, please contact our Customer Service Department.
Cat# Product Name Size 1334 MEGAscript T7 Kit 40 rxns 1626 MEGAscript RNAi Kit 20 rxns 1909 MEGAclear-96 Kit 96 rxns