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Construction and characterization of partial digest DNA libraries made from flow-sorted human chromosome 16
J L Longmire1, N C Brown, L J Meincke
1Life Sciences Division and Center for Human Genome Studies, Los Alamos National Laboratory, NM 87545.
Genetic Analysis, Techniques and Applications
|June 1, 1993
Summary
Researchers developed chromosome-specific partial digest libraries for human chromosome 16. These libraries aid in creating a high-resolution physical map and isolating gene sequences.
Area of Science:
- Genomics
- Molecular Biology
- Cytogenetics
Background:
- High-resolution physical mapping is crucial for understanding genome organization and function.
- Development of specialized libraries is essential for efficient gene mapping and sequencing projects.
- Flow-sorted chromosomes provide a pure source for constructing targeted genomic libraries.
Purpose of the Study:
- To present techniques for creating chromosome-specific partial digest libraries.
- To characterize two such libraries derived from flow-sorted human chromosome 16.
- To provide resources for the National Laboratory Gene Library Project and physical mapping efforts.
Main Methods:
- Flow-sorting of human chromosome 16 from a mouse-human monochromosomal hybrid cell line.
- Construction of partial digest libraries in Charon-40 (LA16NL03) and sCos-1 (LA16NC02) vectors.
- Characterization of library enrichment, nonrecombinant background, and sequence representation.
Main Results:
- Two chromosome 16-specific libraries (LA16NL03 and LA16NC02) were successfully constructed.
- Libraries exhibit approximately 90% enrichment for human chromosome 16 sequences.
- Both libraries demonstrate low nonrecombinant backgrounds and high sequence representation.
Conclusions:
- The developed libraries are valuable resources for physical mapping of human chromosome 16.
- The cosmid library is particularly useful for isolating coding sequences.
- These libraries support ongoing efforts in genomic research and gene mapping.