Buffers that can be used: 10 mM sodium cacodylate-cacodylic acid, 200 mM sodium chloride, 20 mM magnesium chloride at pH 6.8. Typically, annealing is done by heating up to 90 °C, followed by a gradual cooling to room temperature.
More details about how to form triplexes can be reviewed in the following papers:
- Obika, S., Nanbu, D., Hari, Y., Morio, K., In, Y., Ishida, T., Imanishi, T. (1997) Tetrahedron Lett. 38, 873S-8738.
- Torigoe, H., Hari, Y., Sekiguchi, M., Obika, S., Imanishi, T. (2001) J Bio!. Chern. 276, 23S4-2360.
- Rahman, S. M. A., Seki, S., Obika, S., Haitani, S., Miyashita, K., Imanishi, T. (2007) Angew. Chern. Int.Ed., 46, 4306-4309.