| Contributor: |
Simon Dawson at the School of Biomedical Sciences, Queen's Medical Centre
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| Spin column purification can be used to change buffers without a concomitant change in solution volume, to remove protein contaminants, or to purify plasmid DNA suitable for sequencing. If plasmid DNA is required for sequencing purposes, the column should be set up as described, but Sephacryl S-400 is substituted for Sephadex G50. |
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1. Plug a 1 ml syringe barrel with siliconized Glass Wool and fill to the top with a sterile suspension of Sephadex G50 equilibrated with water.
2. Once filled, place the syringe barrel into a 10 ml centrifuge tube and centrifuge at 3000 X g for 5 min in a swinging bucket rotor.
3. Add 100 to 200 μl of the buffer that the DNA is dissolved in is added to the top of the column
4. Centrifuge the column at 3000 X g for 5 min. Repeat this step 3 times.
5. Place the conical base of a sterile microcentrifuge tube into the bottom of the emptied 10 ml centrifuge tube and replace the syringe barrel.
6. Apply the DNA sample to the top of the column in 100 to 200 μl of buffer and centrifuge at 3000 X g for 5 min.
7. Collect the DNA-containing eluate in a microcentrifuge for further manipulation.
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This bioProtocol does not use any solutions
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Sephacryl S-400
Glass Wool
Sephadex G50
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No hints are associated with this bioProtocol
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