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POLY-PAK™ PURIFICATIONThe use of Poly-Pak™ packings in cartridges or barrels overcomes several disadvantages usually associated with reverse phase (RP) cartridges. The packing is stable in the pH range 1-13, thus the ammonium hydroxide solution, diluted with water, is loaded directly onto the packing. Also, after elution of failure sequences, the trityl group is removed and washed from the support-bound oligonucleotide. The fully deprotected product can then be eluted and isolated by lyophilization. Poly-Pak™ Cartridges may also be used for desalting normal or labelled oligonucleotides. The original Poly-Pak cartridge and barrel are designed for 0.2 µmole syntheses or less. Poly-Pak II cartridges and barrels are designed for use with 1 µmole syntheses. A booklet, User Guide To Poly-Pak™ Cartridge Purification, describes in detail the process and several applications. This booklet is delivered with cartridges, barrels, and packing and is available on request. |
Poly-Pak Cartridge Used Manually
Poly-Pak™ is a trademark of Glen Research Corporation |
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Glen-Pak™ CartridgesDMT-on purification of oligonucleotides using traditional cartridges, exemplified by Poly-Pak cartridges, has served the oligo synthesis community well for many years. However, two major deficiencies in the traditional cartridges quickly became apparent: a single loading process did not work, making them problematical for high throughput robotics systems; and the range of successful purification was restricted to around 50mers. The Fluoro-Pak purification system, as applied to oligonucleotide purification, solved both of these problems but at a higher cost. The required fluorous DMT monomers must be available to be added in the final synthesis cycle. Glen-Pak DNA and RNA cartridges have advantages over Poly-Pak cartridges in that a single loading of the diluted crude deprotection solution is all that is necessary. Also, the range of purification has been extended to 100+ using DMT-on oligos. Glen-Pak cartridges have similar performance to Fluoro-Pak cartridges but without the need for the fluorous DMT group at the 5’ terminus, so the cost is lower. In addition, Glen-Pak cartridges allow purification of virtually the complete range of dyes and modifiers. Poly-Pak™ and Glen-Pak™ are trademarks of Glen Research Corporation. Fluoro-Pak™ is a trademark of Berry & Associates, Inc.
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See AlsoPoly-Pak Reagents p116 |
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Fluoro-Pak™ purificationThe fluorous affinity purification of oligonucleotides is a quick and simple affinity-based method for the purification of oligonucleotides that relies on the strong interaction of fluorous-tagged oligonucleotides (made with fluorous-tagged phosphoramidites) with the fluorinated adsorbent present in Fluoro-Pak™ columns. Fluorous affinity purification is operationally similar to DMT-on purification using a reverse-phase (RP) adsorbent, e.g., RP cartridge purification, except that it involves a much stronger affinity interaction and is thus able to afford higher selectivities and recoveries, even with long oligos.
| OTHER INSTRUMENT TYPESAll minor bases, RNA products and modifiers are packaged in septum-capped vials suitable for ABI and other instruments. If you would like another type of vial/column add the following to the end of the catalog number.
INTELLECTUAL PROPERTY“Fluoro-Pak” is a trademark of Berry & Associates, Inc. Products for Fluorous Affinity Purification of Oligonucleotides: Patents applied for, Berry & Associates, Inc. Further, the use of these products is licensed under U.S. Patents 6,673,539, 6,156,896; 5,859,247; and 5,777,121 and one or more pending patents owned or controlled by Fluorous Technologies, Inc. See AlsoPoly-Pak Reagents p116 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Oligo-Affinity SupportOligo-affinity supports (OAS) should ideally be compatible with automated synthesis, should be non-friable, should not shrink or swell, and should have low non-specific binding of the proteins or DNA. On the support shown below is an Adenosine residue attached through the exocyclic amino group. In this way, synthesis progresses regularly on removal of the 5’-DMT group. However, on treatment with ammonium hydroxide, the oligo is not cleaved from the support. This matrix can then be used as an affinity support for a complementary segment of DNA or RNA. Alternatively, the complementary strand can be annealed to the support and the double stranded DNA can be used as an affinity support for purifying DNA binding proteins. We expect that OAS PS will be used for purification of components from biological fluids.
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OTHER INSTRUMENT TYPESAll minor bases, RNA products and modifiers are packaged in septum-capped vials suitable for ABI and other instruments. If you would like another type of vial/column add the following to the end of the catalog number.
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