Glen UnySupport™ is a version of UnyLinker™, shown in Figure 1, that was developed at Isis Pharmaceuticals, and is preferred for high throughput oligonucleotide synthesis. Glen UnySupport is compatible with most deprotection strategies from gas phase deprotection with methylamine to UltraMild deprotection with potassium carbonate in methanol. We supply Glen UnySupport under license from Isis Pharmaceuticals.
Glen UnySupport is fast becoming one of our most popular universal supports since it is compatible with such a wide range of deprotection strategies and conditions, as shown in Table 1.
For instance, Universal Support III (Figure 1) is an excellent universal support for DNA and RNA synthesis and for oligos that require UltraMild conditions because of the presence of sensitive dyes or minor bases. However, since efficient cleavage is dependent upon the cyanoethyl phosphotriester remaining intact until treatment with 2 M NH3 in methanol, it is not compatible with the popular DEA post-synthesis treatment to reduce N3 alkylation by acrylonitrile nor is it compatible with gas phase deprotection conditions.
This is not the case with the Glen UnySupport. We have now established conditions for UltraMild deprotection using both 50 mM Potassium Carbonate in methanol and tert-Butylamine/Water, UltraFast deprotection using AMA, and Gas Phase deprotection using Methylamine. While US III continues to be the universal support of choice for RNA synthesis, in most cases, Glen UnySupport performs as well using a wide variety of popular deprotection cocktails. In Table 1, we have summarized the current cleavage conditions necessary to completely eliminate the Glen UnySupport from the 3’ terminal base.
|Ammonium Hydroxide||2 hours||80°C|
|Ammonium Hydroxide/ 40% Methylamine (AMA)||0.5 hours||80°C|
|Methylamine Gas||0.5 hours||65°C (30 psi)|
|Potassium Carbonate in Anhydrous Methanol||17 hours||Room Temperature|
|t-Butylamine/Water (1:3) v/v||4 hours||60°C|
Universal Supports are the supports of choice for high throughput synthesis since they remove the need for individual wells or columns to contain a unique support. In the past, supports have been added to synthesis plates by slurrying in a dense (and usually toxic) liquid and dispensing conventionally. An alternative approach would be to embed the support into frits suitable for inserting into standard columns or plates.
We now offer high density polyethylene (HDPE) frits with embedded Glen UnySupport for high throughput applications and to allow customers to make their own inexpensive columns. The frits contain 40 nanomoles of Glen UnySupport and fit directly into empty MerMade and AB3900 columns. They can also be used on ABI 394 and Expedite instruments when fitted into inexpensive female-female luer adapters. We are also offering the female-female luers required for use of the Glen UnySupport Frits on Expedite and ABI 394 instruments. To assemble the column, simply place the frit in the luer fitting and seat it in the center using an allen wrench or equivalent tool (be careful not to puncture the frit). The luers are re-useable and can be purchased in packs of 10 (Cat. No. 20-0060-00).