Getting enough full-length oligo?

Coupling efficiency matters!

The coupling efficiency achieved by an oligonucleotide manufacturer has a direct effect on the quality of the oligonucleotides produced. Find out why coupling efficiency should be important to you.

Sep 29, 2017

The coupling efficiency achieved by an oligonucleotide manufacturer has a direct effect on the quality of the oligonucleotides produced. When the coupling efficiency is high, there are fewer truncated products and deletions, resulting in a higher percentage of the desired sequence. In an industry where the standard coupling efficiency is 98.5%, IDT consistently exceeds that. The coupling efficiency for standard oligos from IDT is 99.2% and even higher—99.5%—for Ultramer® Oligonucleotides. This allows us to synthesize 70-base Ultramer oligos and achieve up to 70% full-length product; the same oligo synthesized at an average coupling efficiency of 98.5% by a competing manufacturer might only garner 35% full-length product at most (Figure 1). Thus what may appear as small differences in percentage translates to a dramatic difference in amount of full-length product.


Figure 1. Full-length product synthesized under varying average coupling efficiencies.

A higher coupling efficiency means better performance

Why should coupling efficiency be important to you? Because there are more full length sequences present, you will likely see better and more consistent results in experiments when using an oligonucleotide manufactured with a higher coupling efficiency. This consistency also extends across multiple syntheses of an oligonucleotide. Thus, when re-ordering, you can be confident that the new oligonucleotide will function the same as an oligo from a previous synthesis, contributing to the consistency of your reactions and decreasing efforts needed to troubleshoot anomalous results.

The high coupling efficiency attained by IDT facilitates synthesis of sufficient full-length yields of high quality, long oligos, such as the IDT Ultramer Oligonucleotides. This is one of the reasons IDT is unique in its field, as it is a feat our competitors struggle to accomplish. At the industry standard of 98.5% coupling efficiency, a 150mer is not commercially viable, as only 10% would be full-length.

IDT proprietary synthesis platforms yield the highest quality modified and unmodified oligonucleotides across various lengths. Even when these oligos are ordered without additional purification, the large percentage of full-length product can provide a substantial improvement in the data you collect. Effects of variance in oligonucleotide quality can influence many common molecular techniques, including cloning, mutagenesis, sequencing, and qPCR. While the industry standard is 98.5%, IDT recognizes that even modest gains in coupling efficiency lead to higher quality oligonucleotides and, therefore, improved sensitivity and consistency in experiments.