Probe detection

The method used to detect the regions to which the probe has hybridized to a membrane is dependent on the nature of the label used in generating the probe. Frequently, the label is radioactive (e.g. l2P, "P or "S) and detection is by autoradiography using X-ray film. Detection of single-copy genomic DNA sequences usually requires the use of intensifying screens at low temperatures ( —70°C). Exposure times will depend on many factors and is best determined empirically - it may range from a few minutes to several weeks. Chemiluminescent probes are also detected with X-ray film and, in general, detection times can be shorter than with radiolabeled probes.

A relatively recent advance in detection has been the introduction of the use of electronic imagers which are capable of detecting radioactivity and chemiluminescence as well as direct fluorescence. Their major advantages are increased sensitivity and linearity of response compared with X-ray film.

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