B-pure synthesis protocol

B-pure Oligonucleotide Synthesis protocol produces custom synthesis of affordable, high-quality oligonucleotides up to 300 bases. Oligonucleotides longer than 80 bases are synthesized automatically on this protocol. Our proprietary production and method has improved the quality significantly. Best results have been achieved in numerous experiments which have been performed in our intensive testing program. The B-Pure oligonucleotide synthesis protocol cleans the oligos from any chemicals or unwanted side products. Additional quality control is performed on our state-of-the-art LC-MS system to guarantee highest quality and optimized functionality of the oligonucleotides.

  

Coupling efficiency

Coupling efficiency relates to the final amount of full length sequence after synthesis of your oligonucleotide.  Oligonucleotide synthesis is the step-wise addition of nucleotide residues to the 5'-site of the growing chain until the desired sequence is completed. Each addition of nucleotide residues is called a synthesis cycle. The coupling efficiency defines the efficiency of each synthesis cycle. 

By using the B-Pure synthesis protocol we achieve a coupling efficiency up to 99.9% through an entire 300 base synthesis reaction. A coupling efficiency of 99% or 98% seems to be very good, but on closer examination the full-length yield is only half for a 40 mer! For a 300 mer this will result in a crude yield of less than 5%, which is not acceptable. As demonstrated in the figure below, the only way to synthesize longmers up to 300 bases with an appreciable yield of full length material (>30%) is to have a coupling efficiency of more than 99.5%.

coupling efficiency long oligo synthesis