A DNA probe labeling technique based on the introduction of nucleotide gaps into one strand of double-stranded DNA by a treatment with a nuclease, followed by the use of a DNA polymerase with exonuclease activity to extend the gaps and replace the nucleot
Refers to the in vitro procedure used to radioactively label DNA uniformly to a high specific activity. Nicks are introduced into the unlabelled DNA by an endonuclease, thus creating 3' hydroxyl termini. E. coli DNA polymerase then adds radioactive residues to the 3' hydroxy terminus of the nick; nucleotides are removed from the 5' side. This procedure leads to an identical DNA molecule with the nick located further along the duplex.
A procedure for labelling DNA. A DNA fragment is treated with DNase to produce single-stranded nicks. The nick is moved along the DNA molecule in the presence of labelled deoxyribonucleoside triphosphates by the concerted action of the 5´- 3´ exonuclease and 5´- 3´ polymerase activities of E. coli DNA polymerase I.