»  AMPure Cleanup: Robot or by hand

AMPure Cleanup: Robot or by hand

AMPure Cleanup: Robot or by hand.

1. Transfer 18uL AMPure to an Eppendorf plate.

2. Add 10uL PCR product and mix.

3. Let sit 5 minutes.

4. Place on magnet, let sit 5-10 minutes.

5. Remove entire volume, leaving beads, discard liquid.

6. Add 200uL 70% Ethanol, let sit 30 seconds, flip while still on magnet to remove ethanol.

7. Repeat Ethanol wash.

8. Take plate off magnet; let air dry for 12-25 minutes.

9. Add 40uL H2O, mixing well, to re-suspend the DNA.

10. When you are ready to set up your sequencing reactions,

11. Place plate on magnet for 10 minutes.

 

Faint band concentration:
After AMPure cleanup, place plate on magnet for 10 minutes and transfer product to a new tube or plate. Evaporate to dryness on DNA speedvac. Re-suspend in 10uL H2O and proceed to sequencing reaction.