Chromosomal in situ hybridization with double-labeled DNA: signal amplification at the probe level
Harry Scherthan, Martin Köhler, P. H. Vogt, K. von Malsch, Dieter Schweizer
Abstract
Harry Scherthan, Martin Köhler, P. H. Vogt, K. von Malsch, Dieter Schweizer
Abstract
A double-labeling approach was applied to nonisotopic in situ hybridization with individual cosmid and plasmid clones, using digoxigenin or biotin as label and a combination of two separate enzymatic labeling methods. Probe labeling was achieved by nick translation, followed by tailing of the probe by terminal deoxynucleotidyl transferase. The double-labeling method, in conjunction with an improved detection protocol, provides for a higher signal intensity than that obtainable with single-labeled probes.
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A double-labeling approach was applied to nonisotopic in situ hybridization with individual cosmid and plasmid clones, using digoxigenin or biotin as label and a combination of two separate enzymatic labeling methods. Probe labeling was achieved by nick translation, followed by tailing of the probe by terminal deoxynucleotidyl transferase. The double-labeling method, in conjunction with an improved detection protocol, provides for a higher signal intensity than that obtainable with single-labeled probes.
Key concepts: Nick translation, Digoxigenin, Biology, Molecular biology, Hybridization probe, In situ hybridization, Molecular probe, Cosmid