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High-speed chromosome sorting.
J W Gray1, P N Dean, J C Fuscoe
1Biomedical Sciences Division, Lawrence Livermore National Laboratory, CA 94550.
Science (New York, N.Y.)
|October 16, 1987
Summary
High-speed chromosome sorting using dual-beam technology purifies chromosomes efficiently. This method enables DNA library construction and gene mapping with high purity.
Area of Science:
- Genetics
- Molecular Biology
- Biotechnology
Background:
- Chromosome purification is crucial for genetic research.
- Existing sorting methods have limitations in speed and efficiency.
Purpose of the Study:
- To develop a high-speed dual-beam sorting method for chromosome purification.
- To enable efficient DNA library construction and gene mapping.
Main Methods:
- Utilized dual-beam flow cytometry for chromosome sorting.
- Employed fluorescent dyes Hoechst 33258 and chromomycin A3 for DNA staining.
- Processed human lymphoblast chromosomes at high rates.
Main Results:
- Achieved sorting rates of approximately 200 chromosomes per second.
- Processed up to 20,000 fluorescent objects per second.
- Obtained chromosome purity of about 90 percent for single-type sorting.
- Sorted DNA was suitable for recombinant DNA library construction and gene mapping.
Conclusions:
- Dual-beam high-speed sorting significantly enhances chromosome purification efficiency.
- The developed method provides high-purity chromosomes for downstream genetic applications.
- This technology offers a tenfold increase in sorting speed compared to conventional sorters.