A bacterial artificial chromosome library for sequencing the complete human genome

作者：Osoegawa K; Mammoser AG; Wu CY; Frengen E; Zeng CJ; Catanese JJ; de Jong PJ^*

来源：Genome Research, 2001, 11(3): 483-496.

DOI：10.1101/gr.169601

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.

• 出版日期2001-3