Unit

UNIT 5.2 Construction of YAC Libraries with Large Inserts

  1. Simon Foote1,
  2. Christopher Denny (Mg2+ limitation)2

Published Online: 1 FEB 2002

DOI: 10.1002/0471142905.hg0502s31

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Foote, S. and Denny, C. 2002. Construction of YAC Libraries with Large Inserts. Current Protocols in Human Genetics. 31:5.2:5.2.1–5.2.20.

Author Information

  1. 1

    Whitehead Institute, Cambridge, Massachusetts

  2. 2

    University of California at Los Angeles, School of Medicine, Los Angeles, California

Publication History

  1. Published Online: 1 FEB 2002
  2. Published Print: OCT 2001

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.