Unit

UNIT 25B.1 Production of a Subtracted cDNA Library

  1. Lloyd B. Klickstein

Published Online: 1 AUG 2001

DOI: 10.1002/0471142727.mb25b01s55

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Klickstein, L. B. 2001. Production of a Subtracted cDNA Library. Current Protocols in Molecular Biology. 55:B:25B.1:25B.1.1–25B.1.8.

Author Information

  1. Brigham and Women's Hospital, Boston, Massachusetts

Publication History

  1. Published Online: 1 AUG 2001
  2. Published Print: JUL 2001

Abstract

For some experiments, a complete cDNA library is unnecessary and instead, a subtracted cDNA library is useful. A subtracted cDNA library contains cDNA clones corresponding to mRNAs present in one cell or tissue type and not present in a second type. This cDNA library is used to isolate a set of cDNA clones corresponding to a class of mRNAs, or to aid in the isolation of a cDNA clone corresponding to a particular mRNA where the screening procedure for the cDNA clone is laborious because a specific DNA or antibody probe is unavailable. Since relatively few recombinants are obtained after subtraction, this protocol is for a cDNA library constructed in the λ>10 vector or its equivalent, which allows a high cloning efficiency and permits elimination of nonrecombinants; however, the protocol can be used to produce subtracted cDNA libraries in any vector system.