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- SC 15:45, 19 June 2012 (EDT):
PCR Purification using Invitrogen's PureLink PCR Purification Kit.
- DNA sample of volume V
- PureLink PCR purification column
- Binding buffer
- Wash buffer
- Sterile ddH2O
- Add 4V binding buffer (700 μL for concentrated DNA). Vortex and spin.
- Apply to column. Spin in tabletop centrifuge 10,000 rpm 1 min. Discard flowthrough.
- Apply 650 μL wash buffer to column. Spin 10,000 rpm 1 min. Discard flowthrough.
- Spin max speed 2 min to dry.
- Apply 100 μL sterile ddH2O to column membrane. Take care to not touch membrane with pipet tip.
- Incubate RT 10 min.
- Spin max speed 2 min to elute.
- 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.
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
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