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Title: Microdissection and microcloning of genomic DNA markers from human chromosomal region 11q23

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 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.

Authors:
 [1]; ; ; ;  [2]; ; ; ;  [3]
  1. National Inst. of Radiological Sciences, Chiba (Japan) Kazusa DNA Research Inst., Chiba (Japan)
  2. National Inst. of Radiological Sciences, Chiba (Japan)
  3. Nagasaki Univ. (Japan)
Publication Date:
OSTI Identifier:
6082773
Resource Type:
Journal Article
Journal Name:
Genomics; (United States)
Additional Journal Information:
Journal Volume: 16:1; Journal ID: ISSN 0888-7543
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; DISEASES; GENES; HUMAN CHROMOSOMES; MAPPING; LIBRARIES; SCREENING; DNA HYBRIDIZATION; DNA-CLONING; CHROMOSOMES; CLONING; HYBRIDIZATION; 550400* - Genetics

Citation Formats

Seki, Naohiko, Yamauchi, Masatake, Saito, Toshiyuki, Katakura, Reiko, Hori, Tada-Aki, Ohta, Tohru, Yoshiura, Koh-Ichiro, Jinno, Yoshihiro, and Niikawa, Norio. Microdissection and microcloning of genomic DNA markers from human chromosomal region 11q23. United States: N. p., 1993. Web. doi:10.1006/geno.1993.1154.
Seki, Naohiko, Yamauchi, Masatake, Saito, Toshiyuki, Katakura, Reiko, Hori, Tada-Aki, Ohta, Tohru, Yoshiura, Koh-Ichiro, Jinno, Yoshihiro, & Niikawa, Norio. Microdissection and microcloning of genomic DNA markers from human chromosomal region 11q23. United States. https://doi.org/10.1006/geno.1993.1154
Seki, Naohiko, Yamauchi, Masatake, Saito, Toshiyuki, Katakura, Reiko, Hori, Tada-Aki, Ohta, Tohru, Yoshiura, Koh-Ichiro, Jinno, Yoshihiro, and Niikawa, Norio. 1993. "Microdissection and microcloning of genomic DNA markers from human chromosomal region 11q23". United States. https://doi.org/10.1006/geno.1993.1154.
@article{osti_6082773,
title = {Microdissection and microcloning of genomic DNA markers from human chromosomal region 11q23},
author = {Seki, Naohiko and Yamauchi, Masatake and Saito, Toshiyuki and Katakura, Reiko and Hori, Tada-Aki and Ohta, Tohru and Yoshiura, Koh-Ichiro and Jinno, Yoshihiro and Niikawa, Norio},
abstractNote = {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.},
doi = {10.1006/geno.1993.1154},
url = {https://www.osti.gov/biblio/6082773}, journal = {Genomics; (United States)},
issn = {0888-7543},
number = ,
volume = 16:1,
place = {United States},
year = {Thu Apr 01 00:00:00 EST 1993},
month = {Thu Apr 01 00:00:00 EST 1993}
}