Glen-Pak™ Purification

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 and special phosphoramidites, so the cost is lower. In addition, Glen-Pak cartridges allow purification of virtually the complete range of dyes and modifiers. The Glen-Pak DNA Cartridge 3g is a large cartridge capable of purifying 10-20 µmole oligonucleotide syntheses using the standard DMT-on procedure and Glen-Pak DNA 30mg 96-Well Plates are for parallel purification of up to 50 nmole scale syntheses. Scale suggestions for the Glen-Pak DNA product line are shown below:

Glen-Pak DNA Product	Catalog Number	Synthesis Scale Compatibility
Glen-Pak DNA 50mg Purification Cartridge	60-5000-96	10 nmole – 200 nmole
Glen-Pak DNA Purification Cartridge	60-5100-XX 60-5200-XX	10 nmole – 1.0 µmole
Glen-Pak DNA Cartridge 3G	60-5300-01	5 µmole – 20 µmole
Glen-Pak DNA 30 mg 96-Well Plate	60-5400-01	10 nmole – 50 nmole

Please see our USER GUIDE TO GLEN-PAK™ PURIFICATION

See
Glen Report 19.2
Glen Report 23.2

See Also

Glen Gel-Pak™ DESALTING

The principle of the Glen Research gel filtration column, Glen Gel-Pak™, is based on size exclusion chromatography that separates molecules according to the hydrodynamic volume of the molecule in aqueous solutions. In gel filtration, the mobile phase for size exclusion is an aqueous solution and the stationary phase is a porous resin. The pores of the resin are sized such that they allow small molecules to enter the pores, yet exclude larger molecules from the pores. The small molecules, such as salts and hydrolyzed protecting groups, diffuse into the pores of the resin and move slowly through the column. The larger molecules, such as DNA or proteins, are excluded from the pores and move quickly through the column. The end result is that the larger molecules elute first in the column void volume while the small molecules are still flowing through the resin of the column.

Glen Gel-Pak columns are ideal for desalting and reaction clean up. They can be used for removal of the ammonium hydroxide deprotection solution and hydrolyzed protecting groups after deprotection. The columns can also be used for the clean up of NHS-labelling reactions to separate the labelled oligo and unlabelled oligo from the unreacted NHS ester, the hydrolyzed label, and n-hydroxysuccinimide, thereby greatly simplifying the downstream purification steps.

There are many benefits to Glen Gel-Pak columns: Versatility: Ability to directly desalt oligonucleotides deprotected in either 30% ammonium hydroxide OR 50:50 ammonium hydroxide/40% aqueous methylamine (AMA) Easily exchange buffers Simple clean-up of labelling reactions Mild method for purification from salts and solvents such as DMSO and DMF Capacity: Multiple column sizes (0.2 mL, 1.0 mL and 2.5 mL) are available to match synthesis scale Ability to efficiently desalt short and long oligos at different scales using the same protocol Suitable for oligos >10mer in length

Please see our USER GUIDE TO Glen Gel-Pak™ DESALTING COLUMNS

Oligo-Affinity Support

Oligo-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.