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.
A method of labeling DNA with radioactive or other modified nucleotides by using DNA polymerase I.
a probe labeling technique where DNA polymerase I nicks a starting point from which one strand of a duplex DNA can be degraded and replaced by resynthesis with new labeled precursor nucleotides.
Use of enzymes to break DNA and repolymerize small sections of the molecule, usually for purposes of labeling the DNA with a radioactive nucleotide.
Nick translation is a tagging technique from molecular biology in which endonucleases are used to remove some of the nucleotides of a DNA sequence. Then, Klenow fragment of E. coli DNA polymerase I uses the gaps or nicks as a starting point for replication of DNA. During this elongation process, one or more radioactively or fluorescently labelled nucleotides are used as building blocks for the new DNA synthesized, thus creating a tagged DNA sequence which can be used as a probe in Fluorescent in situ hybridization or blotting techniques.