Reverse synthesis can be achieved by incorporation modifications where the synthesis orientation can be changed as desired. Oligo can be designed for the production of 5'-5' or 3'-3' linkages or a combination of these in the same oligo.
These modified phosphodiester linkage modified oligos are useful in antisense studies, or to synthesize oligonucleotide segments in the opposite sense from normal synthesis, for structural studies.
Having a single inverted base at the 3' position with a 3'-3' linkage imparts the oligo exonuclease resistance and prevents extension by polymerases as there is no free 3' hydroxyl group to initiate synthesis. |