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Vol. 11, Issue 3, 483-496, March 2001

RESOURCES
A Bacterial Artificial Chromosome Library for Sequencing the Complete Human Genome

Kazutoyo Osoegawa,1 Aaron G. Mammoser, Chenyan Wu,2 Eirik Frengen,3 Changjiang Zeng, Joseph J. Catanese,1,2 and Pieter J. de Jong1,2,4

Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, New York 14263, USA

A 30-fold redundant human bacterial artificial chromosome (BAC) library with a large average insert size (178 kb) has been constructed to provide the intermediate substrate for the international genome sequencing effort. The DNA was obtained from a single anonymous volunteer, whose identity was protected through a double-blind donor selection protocol. DNA fragments were generated by partial digestion with EcoRI (library segments 1-4: 24-fold) and MboI (segment 5: sixfold) and cloned into the pBACe3.6 and pTARBAC1 vectors, respectively. The quality of the library was assessed by extensive analysis of 169 clones for rearrangements and artifacts. Eighteen BACs (11%) revealed minor insert rearrangements, and none was chimeric. This BAC library, designated as "RPCI-11," has been used widely as the central resource for insert-end sequencing, clone fingerprinting, high-throughput sequence analysis and as a source of mapped clones for diagnostic and functional studies.

The sequence data described in this paper have been submitted to the GenBank data library under accession nos. AQ936150-AQ936491.]

Present addresses: 1Children's Hospital Oakland Research Institute, 747 Fifty-second Street, Oakland, CA 94609-1809, USA; 2Pfizer Global Research and Development, Alameda Laboratories, 1501 Harbor Bay Parkway, Alameda, CA 94502, USA; 3 The Biotechnology Centre of Oslo, University of Oslo, N-0317 Oslo, Norway.

4 Corresponding author.

11:483-496 ©2001 by Cold Spring Harbor Laboratory Press  ISSN 1088-9051/01 $5.00
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