Unit

UNIT 5.11 Generating Subclones from Large-Insert Genomic Clones

  1. Anthony P. Monaco1,
  2. Zoia Larin2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142905.hg0511s03

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Monaco, A. P. and Larin, Z. 2001. Generating Subclones from Large-Insert Genomic Clones. Current Protocols in Human Genetics. 3:5.11:5.11.1–5.11.6.

Author Information

  1. 1

    John Radcliffe Hospital, Headington, Oxford

  2. 2

    University of Oxford, South Parks Road, Oxford

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: NOV 1994

Abstract

This unit provides a simple protocol for generating a partial-digest sublibrary of yeast DNA containing a YAC of interest. The starting material is high-molecular-weight chromosomal DNA embedded in agarose plugs. Many genome equivalents of these YAC subclones in bacteriophage or cosmid vectors can be plated and screened by hybridization with total human DNA to identify clones that originate from the human portion of the YAC. The human-positive clones can be picked into 96-well microtiter plates for spotting on membranes. Once stored in ordered arrays, the YAC subclones can be constructed into contigs using an end-probe or Alu-PCR hybridization strategy, or a gel fingerprinting technique.This unit provides a simple protocol for generating a partial-digest sublibrary of yeast DNA containing a YAC of interest.