DNA scale of synthesis and typical yeild

Synthesis scale refers to the amount of starting material which is composed solely of the 3’-most nucleotide of a sequence attached to a solid support and housed within the column used to make the oligonucleotide. It is no indication for product yield to be expected! Yield refers to the amount of final product recovered after all of the synthesis and purification steps have been completed.

 Yields of full-length oligonucleotide are lower than the starting amount due to various reasons:

1. The coupling efficiency of different bases or modifications is not 100%. (For example, the standard coupling efficiency of the 4 “normal” DNA bases is about 98-99,5%.) Coupling of modifications can be as low as 50%.
2. Coupling efficiency is influenced by oligo length, oligo sequence and types of modifications. - Bio-Synthesis purifies all oligonucleotides by RP-HPLC. Thus, the yield decreases again, because a majority of unwanted by-products is removed from the crude oligonucleotide.

Bio-Synthesis routinely offers yield guarantees based upon a combination of oligonucleotide length and synthesis scale (please note: yield in our website is based on 20-50 in length for unmodified oligonucleotide). These guarantees are shown in Table 1. Synthesis of very long oligonucleotides (those greater than 60 bases) is problematic. Thus, Bio-Synthesis does not offer any yield guarantees for oligonucleotides greater than 60 bases at our highest synthesis scale and greater than 100 bases at any scale.

 Suggested Reading regarding Synthesis and Yield:

1. Temsamani, J., M. Kubert, and S. Agrawal, Sequence identity of the n-1 product of a synthetic oligonucleotide. Nucleic Acids Res, 1995. 23(11): p. 1841-4.
2. Hecker, K.H. and R.L. Rill, Error analysis of chemically synthesized polynucleotides. Biotechniques, 1998. 24(2): p. 256-60.