Microdissection and Microcloning of Genomic DNA Markers from Human Chromosomal Region 11q23
A human genomic DNA library was constructed by using a microdissection-microcloning procedure with polymerase chain reaction (PCR) techniques on DNA from the chromosome 11q23 region. A total of 450 recombinant pUC clones were isolated from the library. Their insert sizes ranged from 150 to 850 bp wi...
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Published in | Genomics (San Diego, Calif.) Vol. 16; no. 1; pp. 169 - 172 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
San Diego, CA
Elsevier Inc
01.04.1993
Elsevier |
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Abstract | A human genomic DNA library was constructed by using a microdissection-microcloning procedure with polymerase chain reaction (PCR) techniques on DNA from the chromosome 11q23 region. A total of 450 recombinant pUC clones were isolated from the library. Their insert sizes ranged from 150 to 850 bp with a mean of 320 bp. Fifty pUC clones were randomly selected and analyzed in detail. Southern blot analyses showed that 21 (42%) clones were unique DNA sequences, 20 (40%) clones were repetitive sequences, and 9 (18%) clones had no detectable hybridization. The unique sequences were used further in a secondary screening of a partially digested human genomic DNA library constructed in phage vector, and 4 clones were isolated. The chromosomal locations of these phage clones were confirmed to be in the q23 region of chromosome 11 by fluorescence in situ suppression hybridization. These pUC microclones isolated from the chromosomal region-specific genomic DNA library will be useful in the construction of physical contig maps with yeast artificial chromosome and/or cosmid clones and in the positional cloning of disease-associated genes localized to the q23 region of chromosome 11. |
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AbstractList | A human genomic DNA library was constructed by using a microdissection-microcloning procedure with polymerase chain reaction (PCR) techniques on DNA from the chromosome 11q23 region. A total of 450 recombinant pUC clones were isolated from the library. Their insert sizes ranged from 150 to 850 bp with a mean of 320 bp. Fifty clones were randomly selected and analyzed in detail. Southern blot analyses showed that 21 (42%) clones were unique DNA sequences, 20 (40%) clones were repetitive sequences, and 9 (18%) clones had no detectable hybridization. The unique sequences were used further in a secondary screening of a partially digested human genomic DNA library constructed in phage vector, and 4 clones were isolated. The chromosomal locations of these phage clones were confirmed to be in the q23 region of chromosome 11 by fluorescence in situ suppression hybridization. These pUC microclones isolated from the chromosomal region-specific genomic DNA library will be useful in the construction of physical contig maps with yeast artificial chromosome and/or cosmid clones and in the positional cloning of disease-associated genes localized to the q23 region of chromosome 11. 21 refs., 3 figs. A human genomic DNA library was constructed by using a microdissection-microcloning procedure with polymerase chain reaction (PCR) techniques on DNA from the chromosome 11q23 region. A total of 450 recombinant pUC clones were isolated from the library. Their insert sizes ranged from 150 to 850 bp with a mean of 320 bp. Fifty pUC clones were randomly selected and analyzed in detail. Southern blot analyses showed that 21 (42%) clones were unique DNA sequences, 20 (40%) clones were repetitive sequences, and 9 (18%) clones had no detectable hybridization. The unique sequences were used further in a secondary screening of a partially digested human genomic DNA library constructed in phage vector, and 4 clones were isolated. The chromosomal locations of these phage clones were confirmed to be in the q23 region of chromosome 11 by fluorescence in situ suppression hybridization. These pUC microclones isolated from the chromosomal region-specific genomic DNA library will be useful in the construction of physical contig maps with yeast artificial chromosome and/or cosmid clones and in the positional cloning of disease-associated genes localized to the q23 region of chromosome 11. A human genomic DNA library was constructed by using a microdissection-microcloning procedure with polymerase chain reaction (PCR) techniques on DNA from th e chromosome 11q23 region. A total of 450 recombinant pUC clones were isolated from the library. Their insert sizes ranged from 150 to 850 bp with a mean of 320 bp. Fifty pUC clones were randomly selected and analyzed in detail. Southern blot analyses showed that 21 (42%) clones were unique DNA sequences, 20 (40%) clones were repetitive sequences, and 9 (18%) clones had no detectable hybridization. The unique sequences were used further in a secondary screening of a partially digested human genomic DNA library constructed in phage vector, and 4 clones were isolated. The chromosomal locations of these phage clones were confirmed to be in the q23 region of chromosome 11 by fluorescence in situ suppression hybridization. These pUC microclones isolated from the chromosomal region-specific genomic DNA library will be useful in the construction of physical contig maps with yeast artificial chromosome and/or cosmid clones and in the positional cloning of disease-associated genes localized to the q23 region of chromosome 11. |
Author | Yoshiura, Koh-Ichiro Niikawa, Norio Seki, Naohiko Jinno, Yoshihiro Ohta, Tohru Hori, Tada-Aki Katakura, Reiko Yamauchi, Masatake Saito, Toshiyuki |
Author_xml | – sequence: 1 givenname: Naohiko surname: Seki fullname: Seki, Naohiko – sequence: 2 givenname: Masatake surname: Yamauchi fullname: Yamauchi, Masatake – sequence: 3 givenname: Toshiyuki surname: Saito fullname: Saito, Toshiyuki – sequence: 4 givenname: Reiko surname: Katakura fullname: Katakura, Reiko – sequence: 5 givenname: Tohru surname: Ohta fullname: Ohta, Tohru – sequence: 6 givenname: Koh-Ichiro surname: Yoshiura fullname: Yoshiura, Koh-Ichiro – sequence: 7 givenname: Yoshihiro surname: Jinno fullname: Jinno, Yoshihiro – sequence: 8 givenname: Norio surname: Niikawa fullname: Niikawa, Norio – sequence: 9 givenname: Tada-Aki surname: Hori fullname: Hori, Tada-Aki |
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Keywords | Human Genetic marker Microdissection C11-Chromosome Chromosome DNA Molecular cloning Physical map |
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Snippet | A human genomic DNA library was constructed by using a microdissection-microcloning procedure with polymerase chain reaction (PCR) techniques on DNA from the... A human genomic DNA library was constructed by using a microdissection-microcloning procedure with polymerase chain reaction (PCR) techniques on DNA from th e... |
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SubjectTerms | 550400 - Genetics BASIC BIOLOGICAL SCIENCES Biological and medical sciences chromosome 11 Chromosome Mapping CHROMOSOMES Chromosomes, Human, Pair 11 Classical genetics, quantitative genetics, hybrids CLONING Cloning, Molecular DISEASES DNA DNA - genetics DNA HYBRIDIZATION DNA-CLONING Fundamental and applied biological sciences. Psychology Gene Library GENES Genetic Markers Genetics of eukaryotes. Biological and molecular evolution genomes Human HUMAN CHROMOSOMES Humans HYBRIDIZATION In Situ Hybridization, Fluorescence LIBRARIES man MAPPING markers Polymerase Chain Reaction SCREENING |
Title | Microdissection and Microcloning of Genomic DNA Markers from Human Chromosomal Region 11q23 |
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