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Acridine-orange differential fluorescence of fast- and slow-reassociating chromosomal DNA after in situ DNA denaturation and reassociation

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Abstract

A cytological technique based on heat denaturation of in situ chromosomal DNA followed by differential reassociation and staining with acridine orange was developed. Mouse nuclei and chromosomes in fixed cytological preparations show a red-orange fluorescence after thermal DNA denaturation (2–4 minutes at 100° C), and fluoresce green if denaturation is followed by a total DNA reassociation (two minutes or more at 65–66°C). — A reassociation time between a few and 60–90 seconds demonstrates the centromeric heterochromatin of chromosomes (which sometimes aggregate in the form of clusters) and the interphase chromocenters in green, the chromosomal arms fluorescing red-orange. Under the same conditions, the Y chromosome presents a pale green or yellow-green fluorescence along its chromatids, but its centromeric region fluoresces weakly. — The interpretation is suggested that the fast-reassociating chromosomal DNA (as detected by AO in centromeric heterochromatin and interphase chromocenters), represents repetitive DNA.

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Authors and Affiliations

  1. Facultad de Medicina, Centro de Investigaciones en Reproducción, Buenos Aires

    Juan Carlos Stockert & José A. Lisanti

  2. Laboratorio de Genética, Instituto de Oncologia “Angel H. Roffo”, Universidad de Buenos Aires, Argentine

    Juan Carlos Stockert & José A. Lisanti

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  1. Juan Carlos Stockert
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  2. José A. Lisanti
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Stockert, J.C., Lisanti, J.A. Acridine-orange differential fluorescence of fast- and slow-reassociating chromosomal DNA after in situ DNA denaturation and reassociation. Chromosoma 37, 117–130 (1972). https://doi.org/10.1007/BF00284934

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  • DOI: https://doi.org/10.1007/BF00284934

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