A Bacterial Artificial Chromosome Library for Sequencing the Complete Human Genome (original) (raw)
- Kazutoyo Osoegawa1,
- Aaron G. Mammoser,
- Chenyan Wu2,
- Eirik Frengen3,
- Changjiang Zeng,
- Joseph J. Catanese1,2, and
- Pieter J. de Jong1,2,4
- Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, New York 14263, USA
Abstract
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 _Eco_RI (library segments 1–4: 24-fold) and _Mbo_I (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.]
Footnotes
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.
E-MAIL pdejong{at}mail.cho.org; FAX (510) 450-7924.
Article and publication are at www.genome.org/cgi/doi/10.1101/gr.169601.
- Received October 31, 2000.
- Accepted January 9, 2001.
Cold Spring Harbor Laboratory Press