Construction of fifteen human chromosome-specific DNA libraries from flow-purified chromosomes
- 1 January 1986
- journal article
- research article
- Published by S. Karger AG in Cytogenetic and Genome Research
- Vol. 43 (1-2) , 79-86
- https://doi.org/10.1159/000132301
Abstract
We report the construction of 15 human chromosome-specific DNA libraries. Metaphase chromosomes were purified by flow sorting and the DNA was extracted and cleaved with Hindlll before cloning into the λ vector Charon 21 A. A sensitive miniblot hybridization method was used to monitor the physical and biochemical steps in the cloning procedure. Using this method, we have developed a highly efficient protocol for producing large numbers of recombinant phage from 0.2–1.0 × 106 sorted chromosomes. DNA from the following chromosomes was cloned: #4, 6, 8, 9, 11, 13, 14+15, 16, 17, 18, 19, 20, 21, 22 and Y. These libraries are available to the scientific research community and will be valuable in the genetic analysis of the human genome.Keywords
This publication has 14 references indexed in Scilit:
- Correction of a nucleotide-excision-repair mutation by human chromosome 19 in hamster-human hybrid cellsSomatic Cell and Molecular Genetics, 1985
- The DNA‐based human karyotypeCytometry, 1984
- Quantitative karyotyping of human chromosomes by dual beam flow cytometry.Proceedings of the National Academy of Sciences, 1982
- Construction and characterization of genomic libraries from specific human chromosomes.Proceedings of the National Academy of Sciences, 1982
- Cloning of a representative genomic library of the human X chromosome after sorting by flow cytometryNature, 1981
- CONSTRUCTION OF A GENETIC-LINKAGE MAP IN MAN USING RESTRICTION FRAGMENT LENGTH POLYMORPHISMS1980
- Measurement and purification of human chromosomes by flow cytometry and sorting.Proceedings of the National Academy of Sciences, 1979
- Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase IJournal of Molecular Biology, 1977
- Charon Phages: Safer Derivatives of Bacteriophage Lambda for DNA CloningScience, 1977
- A colony bank containing synthetic CoI EI hybrid plasmids representative of the entire E. coli genomeCell, 1976