Corum:Gel Purification

• SC 16:00, 19 June 2012 (EDT):

Overview

Gel Purification using Invitrogen's PureLink Quick Gel Extraction Kit.

Materials

• Gel embedded DNA
• PureLink gel extraction column
• Gel solubilization buffer (L3)
• Isopropynol
• Wash buffer (W1)
• Sterile ddH₂O

Procedure

1. Extract DNA bearing piece of the gel using a razor blade.
2. Weight the gel piece in mg. This number is defined as V.
3. Add 3V gel solubilization buffer. Incubate 55 °C 10 min.
4. Vortex to ensure gel is completely dissolved. If it is not, continue incubating 55 °C and vortexing until complete dissolution.
5. Incubate RT 2 min.
6. Add 1V isopropynol. Vortex and spin.
7. Apply to PureLink gel extraction column. Spin in tabletop centrifuge max speed 1 min. Discard flowthrough.
8. Apply 500 μL wash buffer. Spin max speed 1 min. Discard flowthrough.
9. Spin max speed 2 min to dry.
10. Apply 100 μL sterile ddH₂O to column membrane. Take care to not touch membrane with pipet tip.
11. Incubate RT 10 min.
12. Spin max speed 2 min to elute.
13. 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.