title: Construction of YAC libraries with large inserts.
abstract: The yeast artificial chromosome (YAC) cloning system makes it possible to clone large pieces of genomic DNA into yeast. Libraries have been made containing clones with inserts in the megabase-pair range. The basic protocol in this unit describes preparation of YAC vectors and transformation of ligated DNA into yeast spheroplasts. A support protocol describes titration of Lyticase to make spheroplasts. Additional support protocols detail two methods for partial digestion of genomic DNA: EcoRI restriction endonuclease-EcoRI methylase competition and the partial digestion of genomic DNA by use of limiting amounts of Mg2+, respectively.
authors.affiliation: Whitehead Institute, Cambridge, Massachusetts, USA.
authors.forename: Simon
authors.initials: S
authors.lastname: Foote
authors.forename: Christopher
authors.initials: C
authors.lastname: Denny
mesh_terms: D000818:Animals; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005821:Genetic Techniques; D005822:Genetic Vectors; D005826:Genetics, Medical; D015698:Genomic Library; D006801:Humans; D013104:Spheroplasts / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D004274:DNA, Recombinant