flashPAGE™ Fractionator System
The flashPAGE™ Fractionator System is the fastest, easiest, and most consistent method for PAGE purification of small nucleic acids. Unlike traditional Polyacrylamide Gel Electrophoresis (PAGE), which was designed for the visualization of small nucleic acids, the flashPAGE Fractionator System is designed and optimized for the purification of RNA and DNA.
The innovative flashPAGE Fractionator System enables:
- Ultra-fast PAGE purification of nucleic acids—with standard gel run times of only ~12 minutes
Easy, streamlined protocol
- using ready-to-use reagents, just load your sample and run
- the "lower collection chamber" design eliminates the need for gel excision and elution
- Consistent, reliable results with precise, ready-to-use gels and reagents
Comparison of flashPAGE™ and Traditional PAGE Protocols.
flashPAGE purification is based on passing up to 100 µg nucleic acids through a proprietary denaturing gel matrix, provided as pre-cast gel cartridges, into the lower buffer collection chamber. The lower buffer chamber has been designed for retention of eluted material to facilitate nucleic acid purification. In just a few simple steps, you'll eliminate hours to days of labor (see Figure 1).
Consistent Elution of Nucleic Acids
The flashPAGE A40 Dye Marker, included in the flashPAGE Buffer Kit, elutes at the same molecular weight as a single-stranded RNA molecule of 40 bases. Under normal running conditions, this dye molecule elutes after approximately 12 minutes and can be used as a standard reference point for elution of nucleic acids.
With this system, you can rapidly purify nucleic acids smaller than 40 bases by terminating electrophoresis at the time of dye elution, visible through the instrument's front window. All molecules running through the gel are deposited in the specialized lower collection chamber and can be withdrawn at any point throughout the run. Figure 2 demonstrates the successful isolation of Ambion's Decade™ Markers (RNA molecules between 10–150 nt) using the flashPAGE Fractionator System. In this experiment, two samples were removed, separated by the flashPAGE A40 dye molecule. Following the PAGE run, the flashPAGE Reaction Clean-up Kit is available separately for rapid glass fiber filter (GFF) concentration of nucleic acids and removal of small molecular contaminants.
All components of the flashPAGE Fractionator System are manufactured under strict ISO 9001 requirements. Interested in learning more about this entire system?
Patricia Powers and Rick Conrad • Ambion, Inc.
One of the challenges of miRNA analysis is purification of this fraction of small RNAs, while avoiding other contaminating small RNA species (e.g. tRNA, rRNA, precursor miRNAs, etc.). Since miRNAs are small and typically in low abundance, purification leads to increased sensitivity in miRNA analysis. In addition, accurate microarray profiling requires separation of mature miRNAs from the longer hairpin and pri-miRNA molecules (see Figure 3 at left).
Currently standard polyacrylamide gel electrophoresis (PAGE), followed by gel excision, and nucleic acid elution and concentration, are required for miRNA isolation and purification. This process is time consuming and often imprecise. The flashPAGE™ Fractionator System provides an alternative to PAGE and subsequent gel elution. Mature miRNAs are easily separated from precursor miRNAs and other small RNAs with the help of the flashPAGE™ A40 Dye Marker, which elutes at the same molecular weight as the mature miRNAs. The electrophoresis only takes 12 minutes, and there is no elution step.
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