Enhanced Diagnostic Tools
The addition of a new base to an oligo takes place in four steps:
The difference between the synthesis of a normal oligo with a full phosphodiester backbone and a oligo with a partial or full phophorothioate backbone is the choice of oxidizing agent used in the third step. A phosphodiester bond is produced by using iodine and water to add a fourth oxygen to the phos. A phosphorothioate bond is produced by using Beaucage reagent to add a sulfer to replace the phosphate group. Once either the sulfer or the oxygen has been attached to the phos, the bond is stabilized and will not be affected by the subsequent cycles of chemistry. By switching back and forth between the two oxidizing agents, a chimeric backbone can be constructed.
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