Construction of YAC libraries with large inserts

Current Protocols in Human Genetics
Simon Foote, Christopher Denny

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.

References

Jun 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·H M AlbertsenD Cohen
Nov 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·D R SmithD T Moir
Jun 1, 1987·Analytical Biochemistry·P M Burgers, K J Percival

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