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Studies on locus expansion, library representation, and chromosome walking using an efficient method to screen cosmid
D W Bowden1, H Müller-Kahle, T R Fulton
1Human Genetics Department, Collaborative Research Inc., Waltham, MA 02154.
Gene
|November 30, 1988
Summary
We developed an efficient cosmid library screening method for human genomic DNA. This technique aids in finding polymorphic sequences and simplifies complex genome research, saving labor and time.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Cosmid libraries are crucial for genomic research, but efficient screening methods are needed.
- Identifying homologous cosmids from human genomic DNA aids in locating polymorphic nucleotide sequences.
- Current methods can be labor-intensive, especially for complex genomes.
Purpose of the Study:
- To develop and present an efficient screening method for cosmid libraries.
- To facilitate the isolation of homologous cosmids from human genomic DNA.
- To enable routine cosmid screening for various research applications.
Main Methods:
- Utilized genomic, cDNA, and cosmid probes for screening.
- Applied the method to human chromosomes 7, 10, 16, 17, and X.
- Demonstrated application in chromosome walking experiments at the retinol-binding protein locus.
Main Results:
- Successfully isolated homologous cosmids from multiple human chromosomes.
- The method is applicable to various cosmid cloning systems using restriction enzyme digestion.
- Screening a library of five human genome equivalents requires minimal Southern hybridization filters.
Conclusions:
- The developed method offers an efficient and labor-saving approach to cosmid library screening.
- This technique is valuable for investigating cloned sequence representation and performing chromosome walking.
- The method allows for long-term storage and routine cosmid screening, enhancing laboratory efficiency.