Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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Purification of Labeled Oligonucleotides by Precipitation with Ethanol

(Protocol summary only for purposes of this preview site)

If labeled oligonucleotides are to be used only as probes in hybridization experiments, complete removal of unincorporated label is generally not necessary. However, to reduce background to a minimum, the bulk of the unincorporated label should be separated from the labeled oligonucleotide. Most of the residual unincorporated precursors can be removed from the preparation by differential precipitation with ethanol if the oligonucleotide is >18 nucleotides in length. If complete removal of the unincorporated radiolabel is required (e.g., when the radiolabeled oligonucleotide will be used in primer-extension reactions), chromatographic methods (Protocols 17 and 18) or gel electrophoresis (essentially as described in Chapter 2, Protocol 3) should be used.


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(Limited time special offer.) Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook
Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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