Unit

UNIT 5.5 Screening Large-Insert Libraries by PCR

  1. A.Craig Chinault1,
  2. Nat L. Sternberg (screening P1 libraries)2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142905.hg0505s00

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Chinault, A. and Sternberg, N. L. 2001. Screening Large-Insert Libraries by PCR. Current Protocols in Human Genetics. 00:5.5:5.5.1–5.5.10.

Author Information

  1. 1

    Baylor College of Medicine, Houston, Texas

  2. 2

    Du Pont/Merck Pharmaceuticals, Glenolden, Pennsylvania

Publication History

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

Abstract

This unit describes procedures for screening large-insert libraries by multistep polymerase chain reaction (PCR) analysis of DNA samples from clone pools. In the basic protocol, PCR amplification and agarose gel electrophoresis are used to identify successively smaller pools of DNA or colonies that contain clones with the appropriate-sized amplification product. In the last screening step, individual clones are identified. The first and second support protocols describe the preparation of DNA and yeast-cell pools for screening, and the first alternate protocol describes the preparation of crude lysates suitable for PCR from individual clones or small-scale pools.