PCR Amplification for the Generation of DIG-Labeled Probes

Abstract

The polymerase chain reaction (PCR) is ideally suited to prepare highly specific and efficiently labeled hybridization probes. PCR probes can be strongly reduced in the content of labeled vector sequences if correct strategies for their preparation are applied. They are also defined in length and base pair composition. Thus hybridization events between target and labeled vector sequences can be avoided. Labeled PCR products can be further purified by extraction from agarose gels. However, this is usually not necessary. Cloned templates are to be prefered, but probes could also be generated directly from genomic DNA, if some information on possible primer sequences is available. Labeling DNA by PCR holds advantages over other methods in that the yield of probe is very high, and only a small amount of template DNA is required and the purity of the template DNA is not as crucial as e.g. in random primed labeling.