Size selective DNA precipitation: Difference between revisions
| Line 25: | Line 25: | ||
*Vortex | *Vortex | ||
*Centrifuge 15 min at 10.000 rcf at roomtemperature | *Centrifuge 15 min at 10.000 rcf at roomtemperature | ||
*Carefully remove supernatant not to disturb the pellet, which will be invisible | |||
*Dissolve the pellet in a appropriate amount of buffer of choice | |||
==Critical steps== | ==Critical steps== | ||
Revision as of 15:37, 27 August 2008
Curators
Kersten S. Rabe
Anyone should feel free to add themselves as a curator for this consensus protocol. You do not need to be a curator in order to contribute. The OpenWetWare community is currently discussing the idea of protocol curators. Please contribute.
Abstract
A very fast and easy method for the size-selective removal of smaller DNA from larger fragments. By adjusting the PEG and MgCl2 concentration the range of precipitated DNA fragments can be adjusted.
Materials
Reagents
- DNA to be separated
- 30% (w/v) PEG 8000/30 mM MgCl2 (concentration of PEG 8000 can be varied to shift the size of the percipitated DNA)
- TE Buffer, pH 8.0 (10 mM TRIS-HCl, 1 mM EDTA, pH 8.0)
Equipment
- Centrifuge which can do up to 10.000 rcf (=g)
- Appropriate tubes for the centrifuge
- Pipettes
- Vortexer
Procedure
- Mix 50 μL of sample with 150 µL of TE
- Add 100 µL of PEG/MgCl2
- Vortex
- Centrifuge 15 min at 10.000 rcf at roomtemperature
- Carefully remove supernatant not to disturb the pellet, which will be invisible
- Dissolve the pellet in a appropriate amount of buffer of choice
Critical steps
Troubleshooting
Notes
Acknowledgments
Acnkowledge any help you had in development, testing, writing this protocol.
References
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Specific Protocols
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