27. The reaction mixture of claim 26, wherein the modified nucleoside or nucleotide comprises a locked nucleic acid (LNA), peptide nucleic acid (PNA), 2′-O-methyl (2′-O-Me) modified nucleoside, 2′-fluoro (2′-F) modified nucleoside, and/or a phosphorothioate (PS) bond. 28. The ...
Modelling of a productive P1-substrate complex based on the solved structures suggests steric hindrance as the likely reason for the resistance of Rp-phosphorothioates and phosphorodithioates. Differences with the highly homologous nuclease S1 from Aspergillus oryzae are discussed.Romier CDominguez R...
ii. Oligonucleotide-directed mutagenesis using phosphorothioate substitution. Refer to Appendix A: Site-specific in vitro mutagenesis. iii. Polymerase repair synthesis. Limited Exo III digestion of dsDNA leaves a 3′-OH terminus that can be used as the starting site for primed DNA synthesis by DNA...