Rapid isolation of rare clones from highly complex DNA libraries by PCR analysis of liquid gel pools
Rapid isolation of rare clones from highly complex DNA libraries by PCR analysis of liquid gel pools
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DOI：
10.1016/j.mimet.2006.09.009
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发表时间：
2007-02-01
影响因子：
2.2
通讯作者：
Piel, Joern
中科院分区：
文献类型：
--
作者：
Hrvatin, Sinisa;Piel, Joern
半液体培养基用于筛选高度复杂的 DNA 文库中具有已知序列的克隆的有效方法。在 2 ml 小瓶中生成无偏克隆池，并通过全细胞 PCR 进行筛选，并通过几轮额外的稀释和 PCR 筛选获得单个克隆。为了证明这种方法的实用性，分离了 400,000 名成员宏基因组 fosmid 文库中存在的单个阳性克隆。 (c) 2006 Elsevier B.V. 保留所有权利。
A semiliquid medium was employed in an efficient method to screen highly complex DNA libraries for clones with known sequences. Unbiased clone pools are generated in 2 ml vials and screened by whole-cell PCR, and individual clones are obtained by few additional rounds of dilution and PCR screening. To demonstrate the utility of this approach, the single positive clone present in a 400,000 member metagenomic fosmid library was isolated. (c) 2006 Elsevier B.V. All rights reserved.