A bacterial artificial chromosome library for sequencing the complete human genome

被引：205

作者：

Osoegawa, K ^{[1
]}

Mammoser, AG ^{[1
]}

Wu, CY ^{[1
]}

Frengen, E ^{[1
]}

Zeng, CJ ^{[1
]}

Catanese, JJ ^{[1
]}

de Jong, PJ ^{[1
]}

机构：

[1] Roswell Pk Canc Inst, Dept Canc Genet, Buffalo, NY 14263 USA

来源：

GENOME RESEARCH | 2001年 / 11卷 / 03期

关键词：

D O I：

10.1101/gr.169601

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 a;ingle 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 Mbol (segment 5: sixfold) and cloned into the pBACe3.6 and pTARBACl 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.

引用

页码：483 / 496

页数：14

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