Open AccessIn situhybridisation with fluoresceinated DNA
Author(s) -
J. Wiegant,
Thomas Ried,
Petra M. Nederlof,
M. van der Ploeg,
Hans J. Tanke,
Anton K. Raap
Publication year - 1991
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/19.12.3237
Subject(s) - nick translation , biology , metaphase , microbiology and biotechnology , hybridization probe , dna , in situ hybridization , nucleic acid thermodynamics , nucleic acid , fluorescein , in situ , cytogenetics , molecular probe , fluorescence , genetics , base sequence , chromosome , gene , chemistry , messenger rna , physics , organic chemistry , quantum mechanics
We have used fluorescein-11-dUTP in a nick-translation format to produce fluoresceinated human nucleic acid probes. After in situ hybridization of fluoresceinated DNAs to human metaphase chromosomes, the detection sensitivity was found to be 50-100 kb. The feasibility and the increase in detection sensitivity of microscopic imaging of in situ hybridized, fluoresceinated DNA with an integrating solid state camera for rapid cosmid mapping is illustrated. Combination of fluoresceinated DNA with biotinated and digoxigeninated DNAs allowed easy performance of triple fluorescence in situ hybridization. The potential of these techniques for DNA mapping, cytogenetics and biological dosimetry is briefly discussed.