I have a dsDNA biotinylated on the beads. How can I dissociate the non-biotinylated DNA strand from the biotinylated one?
• There are two methods to dissociate the non-biotinylated DNA from the biotinylated DNA strand. The following protocols are based on using 20 µl of Dynabeads Streptavidin, but are scalable. Both methods may release very small amounts of complementary biotinylated strand from streptavidin. If it is critical that no biotinylated strand is released, either adopt a different biotin modification using dual biotin (two biotin groups in sequence) or covalently bind DNA to e.g. Dynabeads® M-270 Carboxylic Acid. Using heat: • Wash the DNA coated Dynabeads in 50 µl 1 x SSC.* • Resuspend the beads in another 50 µl of 1 x SSC • Incubate at 95 °C for 5 minutes. • Quickly put the tube in magnet stand for 1-2 minutes and transfer the supernatant to a new tube.The supernatant contains your non-biotinylated DNA strand. Using NaOH: • Wash the DNA coated Dynabeads in 50 µl 1 x SSC.* • Resuspend the beads in 20 µl of freshly prepared 0.15 M NaOH. • Incubate at room temperature for 10 minutes. • Put the t
