In situ hybridization with fluoresceinated DNA.
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.
Wiegant
J
J
Department of Cytochemistry and Cytometry, Leiden, The Netherlands.
Ried
T
T
Nederlof
P M
PM
van der Ploeg
M
M
Tanke
H J
HJ
Raap
A K
AK
D015342:DNA Probes / Q000737:chemistry*; D003856:Deoxyuracil Nucleotides / Q000737:chemistry*; D005452:Fluoresceins / Q000737:chemistry*; D006801:Humans; D007150:Immunohistochemistry; D008856:Microscopy, Fluorescence; D009693:Nucleic Acid Hybridization
D015342:DNA Probes; D003856:Deoxyuracil Nucleotides; D005452:Fluoresceins; C069632:fluorescein-11-deoxyuridine triphosphate