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Hi-C: A Method to Study the Three-dimensional Architecture of Genomes.
Published on: May 6, 2010
Human chromosome-specific DNA libraries: construction and purity analysis
J C Fuscoe1, J S McNinch, C C Collins
1Biomedical Sciences Division, Lawrence Livermore National Laboratory, Livermore.
Cytogenetics and Cell Genetics
|January 1, 1989
Summary
Researchers created eight human chromosome-specific DNA libraries using flow-sorted chromosomes. These libraries, containing over five chromosome equivalents each, are now available for scientific use, aiding genetic research.
Area of Science:
- Genomics
- Molecular Biology
- Biotechnology
Background:
- Human chromosome-specific DNA libraries are crucial for genetic research and mapping.
- Previous methods for library construction faced limitations in purity and accessibility.
Purpose of the Study:
- To construct and characterize comprehensive human chromosome-specific DNA libraries.
- To assess and minimize contamination in flow-sorted chromosome libraries.
- To make these valuable resources available to the scientific community.
Main Methods:
- Purification of metaphase chromosomes using flow-sorting.
- DNA extraction, HindIII digestion, and cloning into lambda Charon 21A.
- Quantification of contaminating DNA using species-specific hybridization.
- Assessment of chromosome integrity via fluorescence in situ hybridization (FISH).
Main Results:
- Eight complete human chromosome-specific DNA libraries were generated, each with >5 chromosome equivalents.
- Hamster DNA contamination ranged from 5% to 39%, primarily due to intact hamster chromosomes.
- Less than 0.6% of clones in the chromosome 21 library exhibited a rec+-inhibited phenotype.
Conclusions:
- The developed libraries represent a significant resource for human genetics research.
- Understanding and mitigating contamination sources improves library quality.
- The libraries are accessible through the American Type Culture Collection, facilitating widespread use.
