Construction and characterization of partial digest DNA libraries made from flow-sorted human chromosome 16

Genet Anal Tech Appl. Jun-Aug 1993;10(3-4):69-76. doi: 10.1016/1050-3862(93)90037-j.

Abstract

In this report, we present the techniques used for the construction of chromosome-specific partial digest libraries from flow-sorted chromosomes and the characterization of two such libraries from human chromosome 16. These libraries were constructed to provide materials for use in the development of a high-resolution physical map of human chromosome 16, and as part of a distributive effort on the National Laboratory Gene Library Project. Libraries with 20-fold coverage were made in Charon-40 (LA16NL03) and in sCos-1 (LA16NC02) after chromosome 16 was sorted from a mouse-human monochromosomal hybrid cell line containing a single homologue of human chromosome 16. Both libraries are approximately 90% enriched for human chromosome 16, have low nonrecombinant backgrounds, and are highly representative for human chromosome-16 sequences. The cosmid library in particular has provided a valuable resource for the isolation of coding sequences, and in the ongoing development of a physical map of human chromosome 16.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Chromosomes, Human, Pair 16*
  • Cloning, Molecular
  • Cosmids
  • DNA* / metabolism
  • Deoxyribonucleases, Type II Site-Specific
  • Flow Cytometry
  • Gene Library*
  • Humans
  • Hybrid Cells
  • Mice
  • Phosphorylation

Substances

  • DNA
  • Deoxyribonucleases, Type II Site-Specific
  • GATC-specific type II deoxyribonucleases