Difference between revisions of "Size selective DNA precipitation"
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Revision as of 15:46, 27 August 2008
Kersten S. Rabe
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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.
- 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)
- Centrifuge which can do up to 10.000 rcf (=g)
- Appropriate tubes for the centrifuge
- Mix 50 μL of sample with 150 µL of TE
- Add 100 µL of PEG/MgCl2
- 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
- Before centrifugation mark the tubes in order to know where the pellet will be expected afterwards, as the pellet will be (nearly) invisible
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