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flashPAGE™ Fractionator System
Ultra-Fast Purification of Small Nucleic
Acids
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Figure 1.
The
flashPAGE™ Fractionator System--Ultra Fast Purification
of Small Nucleic Acids. The
flashPAGE Fractionator System consists of the flashPAGE
Fractionator, flashPAGE Pre-cast Gels, and the flashPAGE
Buffer Kit.
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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
Consisting of the flashPAGE Fractionator apparatus, flashPAGE Pre-cast Gels, and flashPAGE Buffers (Figure 1), the flashPAGE Fractionator System provides an efficient alternative to laborious and time-consuming PAGE and subsequent gel elution, which have traditionally been required for small nucleic acid isolation.
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Comparison of flashPAGE™ and Traditional PAGE Protocols. |
Ultra Fast
The flashPAGE System is named to reflect the substantial time and labor savings over traditional PAGE purification of nucleic acids. With it, you will easily and efficiently purify small nucleic acid molecules in run times averaging only 12 minutes.
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 Gels provide consistent separation of molecules, providing predictable separation of small, single-stranded nucleic acids within seconds. Using this system, you can purify RNA or DNA molecules of various sizes using time-based elution and standard controls.
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.
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Figure 2. flashPAGE™ Fractionation. Decade™ Markers (Ambion) in a background of 10 µg mouse brain total RNA were loaded onto a pre-cast flashPAGE Gel Cartridge and electrophoresed with the flashPAGE Fractionator. Two successive fractions were collected. For the first fraction, the lower buffer was collected and precipitated when the dye indicator band had reached the lower end of the gel cartridge. After adding fresh lower buffer to the apparatus, the sample was electrophoresed for 10 more minutes. The second lower buffer fraction was then collected and precipitated. |
High Performance
Each flashPAGE Pre-cast Gel can accommodate up to 100 µg total nucleic acid per column (approximately 10 ng of small RNA is recovered from a 100 µg total RNA sample). Extensive validation of the flashPAGE design confirms that greater than 95% of species longer than 40 nucleotides are excluded from the small RNA/DNA fraction when the run is terminated with the flashPAGE A40 Dye Marker. Faithful representation of RNA is maintained.
Advanced Instrumentation
Although the flashPAGE Fractionator was engineered with the most current design and mechanical circuitry, the apparatus is designed to be an easy addition to any lab. This instrument is provided with a power cord containing standard banana plugs at one end--thus permitting use with most common laboratory power supplies. The flashPAGE Unit to Unit Power Cable makes efficient use of gel power supplies by allowing up to four instruments to run in parallel from one power supply connection.
All components of the flashPAGE Fractionator System are manufactured under strict ISO 9001 requirements. Interested in learning more about this entire system? Contact us directly for answers to your questions (techserv@ambion.com) or visit www.ambion.com/prod/flashPAGE.
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Figure 3. Isolation of Mature microRNA Using flashPAGE™ Fractionator Procedure. |
Scientific Contributors
Patricia Powers and Rick Conrad • Ambion,
Inc.
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