A novel, rapid method for the isolation of terminal sequences from yeast artificial chromosome (YAC) clones
- 1 January 1990
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 18 (10) , 2887-2890
- https://doi.org/10.1093/nar/18.10.2887
Abstract
The recent development of yeast artificial chromosome (YAC) vectors has provided a system for cloning fragments that are over ten times larger than those that can be cloned in more established systems. We have developed a method for the rapid isolation of terminal sequences from YAC clones. The YAC clone is digested with a range of restriction enzymes, a common linker is ligated to the DNA fragments and terminal sequences are amplified using a vector specific primer and a linker specific primer. Sequence data derived from these terminal specific products can be used to design primers for a further round of screening to isolate overlapping clones. The method also provides a convenient method of generating Sequence Tagged Sites for the mapping of complex genomes.Keywords
This publication has 14 references indexed in Scilit:
- A 3.5 genome equivalent multi access YAC library: construction, characterisation, screening and storageNucleic Acids Research, 1990
- A Common Language for Physical Mapping of the Human GenomeScience, 1989
- Alu polymerase chain reaction: a method for rapid isolation of human-specific sequences from complex DNA sources.Proceedings of the National Academy of Sciences, 1989
- Isolation of Single-Copy Human Genes from a Library of Yeast Artificial Chromosome ClonesScience, 1989
- Construction of yeast artificial chromosome libraries with large inserts using fractionation by pulsed-field gel electrophoresisNucleic Acids Research, 1989
- Genome linking with yeast artificial chromosomesNature, 1988
- Diagnosis of α1antitrypsin deficiency by enzymatic amplification of human genomic DNA and direct sequencing of polymerase chain reaction productsNucleic Acids Research, 1988
- A procedure forin vitroamplification of DNA segments that lie outside the boundaries of known sequencesNucleic Acids Research, 1988
- Cloning of Large Segments of Exogenous DNA into Yeast by Means of Artificial Chromosome VectorsScience, 1987
- Toward a physical map of the genome of the nematode Caenorhabditis elegansProceedings of the National Academy of Sciences, 1986