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[A DNA probe suitable for the detection of chromosome 21 copy number in human interphase nuclei by fluorescence in situ hybridization]. 1999

Q Shi, and X Shan, and J Zhang, and X Zhang, and Y Chen, and X Deng, and H Huang, and L Yu, and S Zhao, and Q Zheng, and I Adler
Department of Biology, Nanjing Normal University, Nanjing, Jiangsu, 210097 P. R. China.

OBJECTIVE To prepare DNA probe which can be used to accurately detect chromosome 21 copy number in human interphase nuclei by fluorescence in situ hybridization (FISH). METHODS A probe was produced by universal primer PCR from YAC clone 881D2 and characterized by FISH to metaphases and interphase nuclei of peripheral lymphocytes from 8 normal individuals and 5 patients suffering from trisomy 21. RESULTS A DNA probe was produced and its characteristics were :(1) Most of the probes ranged in size from 350bp to 750bp; (2) The hybridization signals of the probes were located near centromere on long arm of human chromosome 21; (3) In 1524 metaphases scored, about 99.95% of hybridization signals were located on chromosome 21; (4) The signals were very bright and recognizable easily in both metaphases and interphases nuclei; (5) The expected copy number of chromosome 21 was detected by FISH with the probe in more than 98.40% of interphase nuclei and 99.60% of metaphases. CONCLUSIONS The DNA probe reported here is suitable for the detection of chromosome 21 copy number in interphase nuclei and for the study of segregation of chromosome 21 during mitoses of human lymphocytes in culture.

UI MeSH Term Description Entries
D007399 Interphase The interval between two successive CELL DIVISIONS during which the CHROMOSOMES are not individually distinguishable. It is composed of the G phases (G1 PHASE; G0 PHASE; G2 PHASE) and S PHASE (when DNA replication occurs). Interphases
D008297 Male Males
D002467 Cell Nucleus Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed) Cell Nuclei,Nuclei, Cell,Nucleus, Cell
D002891 Chromosomes, Human, Pair 21 A specific pair of GROUP G CHROMOSOMES of the human chromosome classification. Chromosome 21
D005260 Female Females
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D015342 DNA Probes Species- or subspecies-specific DNA (including COMPLEMENTARY DNA; conserved genes, whole chromosomes, or whole genomes) used in hybridization studies in order to identify microorganisms, to measure DNA-DNA homologies, to group subspecies, etc. The DNA probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the DNA probe include the radioisotope labels 32P and 125I and the chemical label biotin. The use of DNA probes provides a specific, sensitive, rapid, and inexpensive replacement for cell culture techniques for diagnosing infections. Chromosomal Probes,DNA Hybridization Probe,DNA Probe,Gene Probes, DNA,Conserved Gene Probes,DNA Hybridization Probes,Whole Chromosomal Probes,Whole Genomic DNA Probes,Chromosomal Probes, Whole,DNA Gene Probes,Gene Probes, Conserved,Hybridization Probe, DNA,Hybridization Probes, DNA,Probe, DNA,Probe, DNA Hybridization,Probes, Chromosomal,Probes, Conserved Gene,Probes, DNA,Probes, DNA Gene,Probes, DNA Hybridization,Probes, Whole Chromosomal
D017404 In Situ Hybridization, Fluorescence A type of IN SITU HYBRIDIZATION in which target sequences are stained with fluorescent dye so their location and size can be determined using fluorescence microscopy. This staining is sufficiently distinct that the hybridization signal can be seen both in metaphase spreads and in interphase nuclei. FISH Technique,Fluorescent in Situ Hybridization,Hybridization in Situ, Fluorescence,FISH Technic,Hybridization in Situ, Fluorescent,In Situ Hybridization, Fluorescent,FISH Technics,FISH Techniques,Technic, FISH,Technics, FISH,Technique, FISH,Techniques, FISH

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