Click-Chemistry based oligonucleotide labeling technology

Currently, labeled and dual labeled oligonucleotide probes are mainly manufactured using either dye-labeled amidites, or activated esters. Both of these methods has its limitations.

For companies and departments involved in custom oligonucleotide synthesis, we offer Click-Chemistry based solution for oligo labeling.

Just like in the case of NHS esters, Click-Chemistry labeling is a two-step protocol. First, alkyne is introduced in oligo using alkyne amidite. Second, deblocked oligo is labeled with azido dye under appropriate conditions. The labeled oligo is then precipitated with acetone, an purified (HPLC is the most appropriate method).

Compared to amidite-based labeling, Click Chemistry yields more pure probes, with higher yield, and the probes require less effort to purify. The sensitive and expensive dye is not subjected to reactive deblocking reagents, and not wasted. There is no need to use ultra-mild protective groups.

Compared to activated ester method, Click Chemistry is more scalable, and requires less dye. The probes are purified more easily. The excess of the dye (about 0.2 to 0.5 eq above required amount) is nearly completely removed during acetone precipitation. This is often not the case for activated ester labeling, where multiple precipitations sometimes required to remove the excessive dye. Our customers use Click Chemistry for the labeling of hundreds of o.u. (thousands of nanomols) of oligonucleotide in a single centrifuge tube.

The cost-efficiency of the labeling method is comparable to amidite and NHS ester labeling. Lumiprobe uses this technology for the production of high quality custom labeled probes containing JOE, cyanines and some of the rhodamines.

We provide all necessary reagents for oligo labeling: alkyne amidites, dye azides, and Click Chemistry catalysts. For your convenience, labeling kits for different dyes are also available.