Proligo® presents the next generation support for reliable, cost-effective oligo synthesis without cross-contamination.
A novel 0.2 μmol ready-to-use Universal ABI 3900 synthesizer column for the synthesis of top quality oligonucleotides has been developed. This new column contains a universal CUTAG loaded CPG 1000 Å frit instead of a polystyrene or CPG support filling, and is therefore powder free. The CPG frit has several advantages over conventional powder filling, which include:
Figure 1.The oligonucleotide synthesis frit consists of a polyethylene-CPG composite material. The CPG is loaded with CUTAG universal anchor, which is related to the Ionis UnyLinker™.1
1 The original UnyLinker™ contains an N-phenyl instead of the N-methyl group, which avoids the release of toxic aniline during the deprotection process. UnyLinker™ is protected by US patent 7,202,264.
This novel column is designed for ABI 3900 synthesizers. It includes the column body and the CUTAG CPG 1000 Å frit. For the synthesis of DNA oligonucleotides there are only a few minor changes compared to the use of nucleoside loaded supports.
Due to the application of the CUTAG universal support, these changes are related to the downstream processing of the synthesized oligonucleotides.
Adjustment of reaction conditions for deprotection
In contrast to nucleoside-loaded supports, the CUTAG support requires dephosphorylation in addition to cleavage and deprotection. If dephosphorylation is incomplete, a modified sequence can be formed as a by-product, whereby the (partial) anchor group remains on the oligonucleotide. To prevent this, we recommend the following deprotection conditions:
Other conditions might be also suitable but should be verified by analysis of the oligonucleotide product by LC-MS.
The novel Proligo® ABI 3900 synthesizer columns are applicable for a number of synthesizer platforms.
Besides the physical properties of the CPG frit, such as the porosity that determines the flow characteristics, the column body design is also critical.
Figure 2.Example of the new ABI 3900 Synthesizer Column with CUTAG CPG Frit
The novel Proligo® ABI 3900 synthesizer column contains a CPG-polymer composite as the solid support. The single piece composite material, or CPG-frit, replaces the three part content of a conventional column, which consists of a bottom and top filter and the CPG or polystyrene powder in between. This modification has several advantages:
Figure 3.AX-HPLC chromatogram of crude dT80 synthesized on a Proligo® 0.2 µmol, ABI 3900 synthesizer column.