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Aug 16, 2018

Version 2

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# AMPure purification protocol V.2

DOI

https://dx.doi.org/10.17504/protocols.io.sk7eczn

Tomasz Suchan¹

¹Institute of Botany, Polish Academy of Sciences

Molecular Biogeography...

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Tomasz Suchan

W. Szafer Institute of Botany, Polish Academy of Sciences

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DOI: https://dx.doi.org/10.17504/protocols.io.sk7eczn

**Protocol Citation:** Tomasz Suchan 2018. AMPure purification protocol. protocols.io
https://dx.doi.org/10.17504/protocols.io.sk7eczn

**License:** This is an open access protocol distributed under the terms of the **Creative Commons Attribution License**, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
**Protocol status:** Working
**correcting some typos**

**Created:** August 14, 2018

**Last Modified:** August 16, 2018

**Protocol Integer ID:** 14719

**Keywords:** ampure purification protocol, purification, protocol

## Guidelines

The protocol below works for 200 µl PCR tubes or plates. If working with 1.5 ml tubes, use larger volumes of EtOH for the washes (step 6 and 7) and Tris/water for the final elution (step 9).

## Troubleshooting
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1. Remove beads from the fridge, equilibrate to the room temperature, mix thoroughly but do not vortex.

2. Add desired ratio of AMPure beads to the DNA to be purified and mix well by pipetting.

3. Incubate 5 minutes.

4. Place on the magnetic rack.

5. Let it stand for 5 minutes on the rack, aspirate and discard supernatant.

6. Add 200 µl of freshly prepared 70% EtOH, incubate 30 seconds, aspirate and discard EtOH.

7. Repeat the wash: add 200 µl of freshly prepared 70% EtOH, incubate 30 seconds, aspirate and discard EtOH.

8. Wait until the pellet dries out completely. DO NOT remove the plate/tubes from the magnetic rack, as the dried AMPure might pop out from the tubes!

9. Add desired volume of Tris 10 mM or water, wetting the dried AMPure pellets (add 1 µl to the final volume to avoid pipetting out the beads)

10. Remove from the magnetic rack.

11. Resuspend by pipetting or vortexing.

12. Incubate 10 minutes, incubating in 37°C can improve DNA yield.

13. Place on the magnetic rack.
14. Let it stand for 5 minutes, pippete out and save supernatant. The eluted DNA is in the supernatant, do not discard it!