Clean Up DNA
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This step uses Illumina Purification Beads to purify the amplified DNA.

• | Illumina Purification Beads (IPB) |
• | Resuspension Buffer (RSB) |
• | Absolute ethanol (EtOH) |
• | Nuclease-free water |

• | IPB |
– | Avoid disturbing the bead pellet when washing. |
– | Use IPB at room temperature. |

1. | Prepare the following consumables: |
• | IPB—Vortex to mix. |
• | RSB—Vortex to mix. |
2. | For each reaction, combine the following volumes to prepare 80% EtOH. |
• | EtOH (400 µl) |
• | Nuclease-free water (100 µl) |
These volumes produce 500 µl 80% EtOH per reaction, including overage.
Vortex to mix.

1. | Remove from the thermal cycler. |
2. | Vortex IPB to resuspend. |
3. | Add 30 μl IPB |
4. | Pipette to mix. |
5. | Incubate at room temperature for 2 minutes. |
6. | Place on the magnetic stand |
7. | Remove and discard all supernatant |
8. | Wash beads as follows. |
a. |
|
b. | Wait 30 seconds. |
c. | Remove and discard all supernatant |
9. | Centrifuge briefly and return to the magnetic stand. |
10. | Using a pipette set to 20 µl, remove all residual EtOH |
11. | Air-dry on the magnetic stand for 2 minutes. |
12. | Remove from the magnetic stand and add 100 μl RSB |
13. | Pipette to mix. |
14. | Incubate at room temperature for 2 minutes. |
15. | Place on the magnetic stand |