Corum:PCR Purification
From OpenWetWare
Jump to navigationJump to search
- 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
- 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.
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
- You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
- Anecdotal observations that might be of use to others can also be posted here.
Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
References
Relevant papers, books, and websites
Contact
or instead, discuss this protocol.