A bacterial artificial chromosome library for sequencing the complete human genome
A bacterial artificial chromosome library for sequencing the complete human genome
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DOI：
10.1101/gr.169601
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发表时间：
2001-03-01
期刊：
影响因子：
7
通讯作者：
de Jong, PJ
中科院分区：
文献类型：
--
作者：
Osoegawa, K;Mammoser, AG;de Jong, PJ
一个30倍冗余的人类细菌人工染色体（BAC）文库与大的平均插入大小（178 kb）已被构建，以提供国际基因组测序工作的中间基板。DNA是从一个匿名的志愿者那里获得的，他的身份通过双盲供体选择协议得到保护。通过用EcoRI（文库片段1-4：24倍）和Mbol（片段5：6倍）部分消化产生DNA片段，并分别克隆到pBACe3.6和pTARBAC 1载体中。通过广泛分析169个克隆的重排和人工产物来评估文库的质量。18个BAC（11%）显示轻微的插入重排，没有嵌合。该BAC文库命名为“RPCI-11”，已被广泛用作插入末端测序、克隆指纹分析、高通量序列分析的中心资源，以及用作诊断和功能研究的定位克隆的来源。
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.