Elsevier

Gene

Volume 49, Issue 2, 1986, Pages 263-271
Gene

Factors which equalize the representation of genome segments in recombinant libraries

https://doi.org/10.1016/0378-1119(86)90287-8Get rights and content

Abstract

Genomic segments which contain inverted repetitions longer than 300 bp are frequently lost from recombinant libraries grown on rec+ hosts. We have found that 9% of phage A clones that contain 15–20-kb insertions of human or Drosophila DNA are inhibited on rec+ hosts and as a result will become underrepresented in amplified genomic libraries. We have therefore examined several factors of both host and vector origin which affect the fidelity of representation of genomic sequences in recombinant DNA libraries constructed in bacteriophage λ vectors. This loss may be diminished if the vector carries either a chi element or a functional gam gene. The most successful approach, however, involves using a host with mutations in recB, recC, and sbcB, or in recD. We have shown that recombinant clones which require such mutant hosts for growth are somewhat more likely to contain DNA derived from loci in the genome which are polymorphic than are clones recovered on conventional hosts.

References (26)

  • D. Barker et al.

    A locus on chromosome 1 Ip with multiple restriction site polymorphisms

    Am.I. Hum. Genet.

    (1984)
  • D. Botstein et al.

    Construction of a genetic linkage map in man using restriction fragment length polymorphism

    Am. J. Hum. Genet.

    (1980)
  • J. Braman et al.

    Highly polymorphic DNA markers useful for the construction of a linkage map of the human genome

    Am. J. Hum. Genet.

    (1985)
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