Molecular cytogenetics using fluorescence in situ hybridization

Conference · Fri Dec 07 00:00:00 EST 1990

OSTI ID:6229656

Gray, J W; Kuo, Wen-Lin; Lucas, J; Pinkel, D; Weier, H-U; Yu, Loh-Chung

Fluorescence in situ hybridization (FISH) with chromosome-specific probes enables several new areas of cytogenetic investigation by allowing visual determination of the presence and normality of specific genetic sequences in single metaphase or interphase cells. in this approach, termed molecular cytogenetics, the genetic loci to be analyzed are made microscopically visible in single cells using in situ hybridization with nucleic acid probes specific to these loci. To accomplish this, the DNA in the target cells is made single stranded by thermal denaturation and incubated with single-stranded, chemically modified probe under conditions where the probe will anneal only with DNA sequences to which it has high DNA sequence homology. The bound probe is then made visible by treatment with a fluorescent reagent such as fluorescein that binds to the chemical modification carried by the probe. The DNA to which the probe does not bind is made visible by staining with a dye such as propidium iodide that fluoresces at a wavelength different from that of the reagent used for probe visualization. We show in this report that probes are now available that make this technique useful for biological dosimetry, prenatal diagnosis and cancer biology. 31 refs., 3 figs.

• Research Organization: Lawrence Livermore National Lab., CA (USA)
• Sponsoring Organization: PUBHEAL
• DOE Contract Number: W-7405-ENG-48
• OSTI ID: 6229656
• Report Number(s): UCRL-JC-105766; CONF-9007183-4; ON: DE91007340; CNN: HD17665; CA45919
• Resource Relation: Conference: Clinical applications of flow cytometry and image analysis, Iwate (Japan), 1-3 Jul 1990
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CHROMOSOMES
GENETIC MAPPING
DNA
PROBES
DNA HYBRIDIZATION
DIAGNOSTIC USES
BIOLOGICAL DOSEMETERS
CARCINOGENESIS
CYTOLOGICAL TECHNIQUES
DNA SEQUENCING
DNA-CLONING
FLUORESCENCE
MOLECULAR BIOLOGY
CLONING
DOSEMETERS
HYBRIDIZATION
LUMINESCENCE
MAPPING
MEASURING INSTRUMENTS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PATHOGENESIS
STRUCTURAL CHEMICAL ANALYSIS
USES
550400* - Genetics
550300 - Cytology
550200 - Biochemistry