Unit

UNIT 5.6 Ligation of Linkers or Adapters to Double-Stranded cDNA

  1. Lloyd B. Klickstein1,
  2. Rachael L. Neve2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb0506s13

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Klickstein, L. B. and Neve, R. L. 2001. Ligation of Linkers or Adapters to Double-Stranded cDNA. Current Protocols in Molecular Biology. 13:III:5.6:5.6.1–5.6.10.

Author Information

  1. 1

    Brigham and Women's Hospital, Belmont, Massachusetts

  2. 2

    McLean Hospital, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 1991

Abstract

The Basic Protocol describes the strategy of producing a genomic DNA library. A number of small-scale ligations are performed using a set amount of vector and varying amounts of insert. Test ligations are transformed into bacteria (plasmid vectors) or packaged and plated on host bacteria (λ and cosmid vectors). The number of clones in the different ligations is compared, and the optimum ratio of vector to insert is indicated by the ligation with the most recombinant clones. A large-scale ligation is then set up using this optimum ratio. This protocol employs a bacteriophage vector; however, cosmid or plasmid vectors can be used with minor modifications.