A bacterial artificial chromosome library for sequencing the complete human genome.
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.
Osoegawa
K
K
Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
Mammoser
A G
AG
Wu
C
C
Frengen
E
E
Zeng
C
C
Catanese
J J
JJ
de Jong
P J
PJ
D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D020451:Contig Mapping; D004276:DNA, Satellite / Q000235:genetics; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D015321:Gene Rearrangement; D005819:Genetic Markers / Q000235:genetics; D015698:Genomic Library; D016045:Human Genome Project; D006801:Humans; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA / Q000379:methods*; D016324:Sequence Tagged Sites
D004276:DNA, Satellite; D005819:Genetic Markers; D011993:Recombinant Fusion Proteins; D015246:Deoxyribonuclease EcoRI; D015252:Deoxyribonucleases, Type II Site-Specific; C024405:GATC-specific type II deoxyribonucleases