Unit

UNIT 6.2 Plating and Transferring Cosmid and Plasmid Libraries

  1. John H. Weis

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb0602s24

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Weis, J. H. 2001. Plating and Transferring Cosmid and Plasmid Libraries. Current Protocols in Molecular Biology. 68:I:6.2:6.2.1–6.2.3.

Author Information

  1. Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: OCT 1993

Abstract

There are two commonly used protocols for the screening of recombinant bacteria with hybridization probes. One method involves the spreading of bacteria on the surface of agar using a sterile spreader. A nitrocellulose membrane filter is then placed on top of the colonies and most of each colony is transferred to the filter. This method works well when relatively small numbers of positive colonies are being selected (up to several thousand). A second method, described in this unit, employs a matrix of some type (here nitrocellulose filters are used) upon which bacteria can be plated and grown when the filter is placed on top of a nutrient agar surface. Once the plated bacteria have grown into visible colonies, the filters can be used for replica plating and in situ hybridization analysis.