Unit

UNIT 5.9 Deriving Probes From Large-Insert Clones by PCR Methods

  1. Hans Albertsen1,
  2. Andrew Thliveris (inverse PCR)1,
  3. John H. Riley (bubble-vector PCR)2,
  4. David J. Munroe (IRS-vector PCR, inter-IRS PCR, IRS-bubble PCR, T vector construction, polishing Alu-PCR products)3,
  5. Paul Watkins (total yeast DNA, Alu PCR from YACs for FISH, scrambling Alu elements)4,
  6. Craig T. Basson (degenerate primer-vector PCR)5

Published Online: 1 MAY 2001

DOI: 10.1002/0471142905.hg0509s17

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Albertsen, H., Thliveris, A., Riley, J. H., Munroe, D. J., Watkins, P. and Basson, C. T. 2001. Deriving Probes From Large-Insert Clones by PCR Methods. Current Protocols in Human Genetics. 17:5.9:5.9.1–5.9.37.

Author Information

  1. 1

    University of Utah, Salt Lake City, Utah

  2. 2

    Glaxo Wellcome Medicines Research Centre, Herts, United Kingdom

  3. 3

    Sequana Therapeutics, La Jolla, California

  4. 4

    Sequana Therapeutics, La Jolla, California

  5. 5

    Cornell University Medical College, New York, New York

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: MAY 1998

Abstract

This unit describes several polymerase chain reaction (PCR)-based methods to obtain DNA fragments from clones with large inserts without prior knowledge of the insert DNA sequence. The protocols can be categorized into three groups: (1) methods to generate DNA fragments at random representing the entire length of the cloned insert, (2) methods to generate DNA fragments representing the extremities of an insert, and (3) methods to generate complex probes suitable for fluorescence in situ hybridization. Support protocols describe direct cloning of these PCR products and the isolation of total yeast DNA from yeast artificial chromosome (YAC) clones.