Hybrid capture Sample Clauses

Hybrid capture. The PCR plate was kept at 65 oC while the hybridization mixture was added from the thermal cycler to the bead solution and the plate was sealed and mixed by inverting it x5 times. Minimal evaporation was essential. The hybrid-capture/bead solution was incubated on a heat block for 30 minute at room temperature and the mix was briefly spanned in a centrifuge. The beads and buffer were separated on the Dynal magnetic separator and the supernatant was removed. The beads were re-suspended in 500 μl of SureSelect Wash Buffer #1 and vortexed for 5 seconds. The samples were incubated for 15 minutes at room temperature and occasionally vortexed. Beads and buffer were again separated on the Dynal magnetic separator and the supernatant was removed. The beads were re-suspended in 500 μl of 65 oC prewarmed SureSelect Wash Buffer #2 and mixed on a vortex for 5 seconds. The samples were incubated for 10 minutes at 65 oC and briefly spun in a centrifuge (<3 seconds). The beads and buffer were separated on the Dynal magnetic separator and the supernatant was removed over a total of 3 washes. The beads were mixed in 50 μl of SureSelect Elution Buffer on a vortex mixer for 5 seconds and samples were incubated at room temperature for 10 minutes. The beads and buffer were separated on the Dynal magnetic separator and the supernatant containing the captured DNA was transferred to a new 1.5 ml microfuge tube. 50 μl of SureSelect Neutralization Buffer were added to the captured DNA.