Size selective DNA precipitation

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(Equipment)
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==Procedure==
==Procedure==
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*Mix 50 μL of sample with 150 µL of TE
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*Add 100 µL of PEG/MgCl<sub>2</sub>
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*Vortex
==Critical steps==
==Critical steps==

Revision as of 17:35, 27 August 2008

Contents

Curators

Kersten S. Rabe

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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

Procedure

  • Mix 50 μL of sample with 150 µL of TE
  • Add 100 µL of PEG/MgCl2
  • Vortex

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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Discussion

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