Why is purification required in the manufacturing of synthetic oligonucleotides?
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چکیده
In order to assemble the sequence, addition of each nucleoside requires four chemical reaction steps and each of these reaction steps is expected to be quantitative (100%). In reality, efficiencies of these reaction steps vary from 97 to 99.9% depending on the reagents, nucleoside building blocks (phosphoramidite) and moisture in the solvent. Depending on the efficiency of each nucleoside addition (four reaction steps), oligonucleotide sequences of twenty nucleosides (20-mer) and twenty five nucleosides (25-mer), theoretically would result in the following percentage of desired oligonucleotide (as shown in the table).
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