AMPure Mods

DNA seq 0.7 Eliminates the 100 and 200bp bands, retains > 300bp bands RNA seq 0.8 Eliminates the 100 band, retains > 200bp bands For everyday cleaning 1.5

DNA seq AMPure Sample+Water 45 µl 65 µl 90 µl 130 µl

RNA seq AMPure Sample+Water 45 µl 55 µl 90 µl 110 µl

1. Gently shake AMPure bottle to resuspend magnetic particles that have settled. 2. Add the desired volume of Agencourt AMPure beads 3. Incubate samples+water + AMPure at room temperature for 5 minutes. 4. Magnetize and then incubate for 5 minutes. 5. Aspirate the cleared supernatant and discard. 6. Dispense 200 µl of 80% EtOH into each tube and incubate for 30 seconds. Aspirate the EtOH wash and discard. Repeat for a total of two washes. 7. Remove all traces of EtOH with a fine pipette tip, and place on bench to air dry for 10 minutes. 8. Add the desired volume of elution buffer (10 mM Tris-HCl, pH 8.0 or water) and vortex for 15-30 seconds. 9. Incubate the samples at room temperature for 5 minutes. 10. Magnetize the sample one last time for 5 minutes. 11. Aspirate the supernatant (the ELUTED DNA) and transfer to a new tube.

TIPS!!! Don’t let the beads go dry between wash steps. Don’t over try beads in the spin-vac or PCR machine; air-temp drying is preferred.