Corum:PCR Purification

• SC 15:45, 19 June 2012 (EDT):

Overview

PCR Purification using Invitrogen's PureLink PCR Purification Kit.

Materials

• DNA sample of volume V
• PureLink PCR purification column
• Binding buffer
• Wash buffer
• Sterile ddH₂O

Procedure

1. Add 4V binding buffer (700 μL for concentrated DNA). Vortex and spin.
2. Apply to column. Spin in tabletop centrifuge 10,000 rpm 1 min. Discard flowthrough.
3. Apply 650 μL wash buffer to column. Spin 10,000 rpm 1 min. Discard flowthrough.
4. Spin max speed 2 min to dry.
5. Apply 100 μL sterile ddH₂O to column membrane. Take care to not touch membrane with pipet tip.
6. Incubate RT 10 min.
7. Spin max speed 2 min to elute.
8. Optional: Combine like samples and/or vacuum centrifuge at 60 °C to reduce volume. IMPORTANT: Time depends on initial and desired final volumes; samples should be checked frequently to prevent total evaporation.

Notes

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References

Relevant papers, books, and websites

1. Invitrogen Life Technologies

Contact

• Sean P Corum

or instead, discuss this protocol.