An anchored framework BAC map of mouse chromosome 11 assembled using multiplex oligonucleotide hybridization
An anchored framework BAC map of mouse chromosome 11 assembled using multiplex oligonucleotide hybridization
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DOI：
10.1006/geno.1998.5620
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发表时间：
1998-12-15
期刊：
影响因子：
4.4
通讯作者：
Bradley, A
中科院分区：
文献类型：
--
作者：
Cai, WW;Reneker, J;Bradley, A
尽管许多生物体的图书馆资源丰富，但这些生物体的物理图谱由于缺乏而受到严重限制。高效的文库筛选技术。我们开发了一种基于高密度基因组过滤器上的多重寡核苷酸杂交的大规模基因组文库筛选的高效策略。我们应用这种策略生成了小鼠 ii 号染色体的细菌人工染色体 (BAC) 锚定图。利用麻省理工学院小鼠 SSLP 数据，通过“overgo”计算机程序设计了 320 对寡核苷酸探针，该程序选择新的引物序列以避免微卫星重复。这些探针识别的 BAC 会自动锚定到染色体上。 92% 的探针从 5.9 倍覆盖的小鼠 BAC 文库中鉴定出阳性克隆，每个标记平均有 7 个阳性克隆。通过 PCR 平均确认了 4.2 个克隆的 204 个标记。我们的数据表明，使用这种策略可以以最少的成本从大型基因组文库中有效地分离出大量克隆。努力。该策略将广泛应用于人类和其他大型基因组的大规模绘图和测序。 (C) 1998 年学术出版社。
Despite abundant library resources for many organisms, physical mapping of these organisms has been seriously limited due to lack. of efficient library screening techniques. We have developed a highly efficient strategy for large-scale screening of genomic libraries based on multiplex oligonucleotide hybridization on high-density genomic filters. We have applied this strategy to generate a bacterial artificial chromosome (BAC) anchored map of mouse chromosome ii. Using the MIT mouse SSLP data, 320 pairs of oligonucleotide probes were designed with an "overgo" computer program that selects new primer sequences that avoid the microsatellite repeat. BACs identified by these probes are automatically anchored to the chromosome. Ninety-two percent of the probes identified positive clones from a 5.9-fold coverage mouse BAC library with an average of 7 positive clones per marker. An average of 4.2 clones was confirmed for 204 markers by PCR. Our data show that a large number of clones can be efficiently isolated from a large genomic library using this strategy with minimal. effort. This strategy will have wide application for large-scale mapping and sequencing of human and other large genomes. (C) 1998 Academic Press.