title: Genomic libraries: I. Construction and screening of fosmid genomic libraries.
abstract: Large insert genome libraries have been a core resource required to sequence genomes, analyze haplotypes, and aid gene discovery. While next generation sequencing technologies are revolutionizing the field of genomics, traditional genome libraries will still be required for accurate genome assembly. Their utility is also being extended to functional studies for understanding DNA regulatory elements. Here, we present a detailed method for constructing genomic fosmid libraries, testing for common contaminants, gridding the library to nylon membranes, then hybridizing the library membranes with a radiolabeled probe to identify corresponding genomic clones. While this chapter focuses on fosmid libraries, many of these steps can also be applied to bacterial artificial chromosome libraries.
authors: Sequencing Research and Development, Wellcome Trust Sanger Institute, Cambridge, UK.
authors: Mike A
authors: MA
authors: Quail
authors: Lucy
authors: L
authors: Matthews
authors: Sarah
authors: S
authors: Sims
authors: Christine
authors: C
authors: Lloyd
authors: Helen
authors: H
authors: Beasley
authors: Simon W
authors: SW
authors: Baxter
mesh_terms: D001435:Bacteriophages; D002999:Clone Cells; D004247:DNA / Q000235:genetics*; D059018:DNA Contamination; D015342:DNA Probes / Q000378:metabolism; D004560:Electricity; D004926:Escherichia coli / Q000821:virology; D005822:Genetic Vectors / Q000235:genetics; D015698:Genomic Library; D023281:Genomics / Q000379:methods*; D008567:Membranes, Artificial; D015203:Reproducibility of Results
chemical_list: D015342:DNA Probes; D008567:Membranes, Artificial; D004247:DNA