Non-radioactive techniques for the labelling of nucleic acids.
Non-radioactively labelled probes potentially have several advantages over radioactively labelled ones, such as increased stability and reduced hazard. As yet, no non-radioactive methods of labelling are as robust or produce as sensitive a probe as 32P. However, there are many options, some of which use approaches familiar to 32P users (nick translation, random priming, tailing). The majority of methods, whether enzymatic or chemical, direct or indirect, ultimately require the detection of an enzyme by a colourimetric or chemiluminescent substrate. Such detection can be achieved variously by direct visualisation (e.g. of a colourimetrically stained blot), by film, by microtitre plate reader or by CCD camera, depending on the choice of assay format and the degree of quantification required.
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