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Related Experiment Videos

A human chromosome 11 NotI end clone library

J Sanford1, B W Kim, L L Deaven

  • 1Department of Human Genetics, Roswell Park Cancer Institute, New York State Department of Health, Buffalo 14263.

Genomics
|March 1, 1993
PubMed
Summary

Researchers created a novel NotI end clone library for human chromosome 11 physical mapping. This library aids in identifying previously unrecognized DNA fragments and understanding gene expression on chromosome 11.

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Area of Science:

  • Genomics
  • Molecular Biology
  • Human Genetics

Background:

  • Construction of a NotI end clone library from a human-hamster hybrid cell line harboring human chromosome 11.
  • Characterization of 51 selected NotI clones to assess library utility.

Purpose of the Study:

  • To develop a valuable resource for physical mapping of human chromosome 11.
  • To identify novel DNA fragments and analyze gene expression patterns on chromosome 11.

Main Methods:

  • Utilizing NotI end clones to isolate NotI linking cosmids.
  • Employing pulsed-field gel electrophoresis for adjacent NotI fragment identification.
  • Mapping clones via fluorescence in situ hybridization and a somatic cell hybrid panel.
  • Analyzing transcript detection using Northern blot analysis.

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Main Results:

  • The majority of NotI clones hybridize to 15- to 200-kb fragments, revealing previously unrecognized fragments.
  • NotI end clones facilitated the identification of adjacent NotI fragments on pulsed-field gels.
  • Clone localization across chromosome 11 showed a nonrandom distribution.
  • Northern blot analysis revealed that 57% of examined NotI clones detected poly(A)+ transcripts in HeLa cell RNA.

Conclusions:

  • The NotI end clone library is a valuable tool for high-resolution physical mapping of human chromosome 11.
  • The library aids in discovering novel genomic regions and understanding gene expression.
  • Observed nonrandom distribution suggests potential regulatory mechanisms or structural features on chromosome 11.