The construction and analysis of M13 libraries prepared from YAC DNA
Open Access
- 25 February 1995
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 23 (4), 670-674
- https://doi.org/10.1093/nar/23.4.670
Abstract
Yeast artificial chromosomes (YACs) provide a powerful way to isolate and map large regions of genomlc DNA and their use In genome analysis is now extensive. We modified a series of procedures to produce high quality shotgun libraries from small amounts of YAC DNA. Clones from several different libraries have been sequenced and analyzed for distribution, sequence integrity and degree of contamination from yeast DNA. We describe these procedures and analyses and show that sequencing at about 1-fold coverage, followed by database comparison (survey sequencing) offers a relatively quick method to determine the nature of previously uncharacterized cosmid or YAC clones.Keywords
This publication has 17 references indexed in Scilit:
- A survey of expressed genes in Caenorhabditis elegansNature Genetics, 1992
- The human HPRT gene on a yeast artificial chromosome is functional when transferred to mouse cells by cell fusionGenomics, 1991
- Basic local alignment search toolJournal of Molecular Biology, 1990
- Low pressure DNA shearing: a method for random DNA sequence analysisNucleic Acids Research, 1990
- Genome linking with yeast artificial chromosomesNature, 1988
- Cloning of Large Segments of Exogenous DNA into Yeast by Means of Artificial Chromosome VectorsScience, 1987
- [7] Random cloning and sequencing by the M13/dideoxynucleotide chain termination methodMethods in Enzymology, 1987
- Separation of Large DNA Molecules by Contour-Clamped Homogeneous Electric FieldsScience, 1986
- Toward a physical map of the genome of the nematode Caenorhabditis elegansProceedings of the National Academy of Sciences, 1986
- Random subcloning of sonicated DNA: Application to shotgun DNA sequence analysisAnalytical Biochemistry, 1983