Deoxyribonucleic acid sequence mapping on metaphase chromosomes by immunoelectron microscopy
Journal Article
·
· Scanning Microscopy, Supplement; (USA)
OSTI ID:7026969
- Univ. of California, Irvine (USA)
Nucleic acid sequences can be localized on chromosomes in the electron microscope after hybridization with a biotinylated DNA probe followed by detection with a primary antibiotin antibody and a secondary antibody coupled to colloidal gold. Hybridization probes can also be labelled with alternative ligands such as N-acetoxy-2-acetylaminofluorene (AAF), Dinitrophenyl-dUTP and Digoxigenin-dUTP. Multiple labelling is possible if these differently modified DNA probes are used in conjunction with colloidal gold preparations of varying particle sizes. A substantial signal amplification can be achieved by incubating preparations with successive cycles of primary antibiotin antibody followed by a biotinylated secondary antibody. Detection is with Streptavidin-gold, and in the case of highly and moderately repeated sequences, the signal is visible in the light microscope. Detailed protocols are given for EM in-situ hybridization to whole mount metaphase chromosomes and include instructions necessary to perform multiple sequence localization and signal amplification.
- OSTI ID:
- 7026969
- Journal Information:
- Scanning Microscopy, Supplement; (USA), Journal Name: Scanning Microscopy, Supplement; (USA) Vol. 3; ISSN 0892-953X; ISSN SMSUE
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201 -- Biochemistry-- Tracer Techniques
550401* -- Genetics-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
ANTIBODIES
CELL CYCLE
CHROMOSOMES
COLLOIDS
DISPERSIONS
DNA
DNA HYBRIDIZATION
DNA SEQUENCING
ELECTRON MICROSCOPY
GENE AMPLIFICATION
GENETIC MAPPING
GOLD ISOTOPES
HYBRIDIZATION
ISOTOPE APPLICATIONS
ISOTOPES
LIGANDS
MAMMALS
MAN
MAPPING
MICE
MICROSCOPY
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PRIMATES
RABBITS
RADIOASSAY
RODENTS
STRUCTURAL CHEMICAL ANALYSIS
TRACER TECHNIQUES
VERTEBRATES
550401* -- Genetics-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
ANTIBODIES
CELL CYCLE
CHROMOSOMES
COLLOIDS
DISPERSIONS
DNA
DNA HYBRIDIZATION
DNA SEQUENCING
ELECTRON MICROSCOPY
GENE AMPLIFICATION
GENETIC MAPPING
GOLD ISOTOPES
HYBRIDIZATION
ISOTOPE APPLICATIONS
ISOTOPES
LIGANDS
MAMMALS
MAN
MAPPING
MICE
MICROSCOPY
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PRIMATES
RABBITS
RADIOASSAY
RODENTS
STRUCTURAL CHEMICAL ANALYSIS
TRACER TECHNIQUES
VERTEBRATES