Multiplexed programmable release of captured DNA.
Multiplexed programmable release of captured DNA.
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DOI：
10.1002/cbic.201402343
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发表时间：
2014-11-03
期刊：
影响因子：
3.2
通讯作者：
Smith, Lloyd M.
中科院分区：
文献类型：
--
作者：
Kennedy-Darling, Julia;Holden, Matthew T.;Shortreed, Michael R.;Smith, Lloyd M.
核酸杂交被广泛用于从复杂样品中特异性捕获互补序列。它适用于两种分析方法，如阵列杂交（如转录组分析，遗传变异分析）和制备策略，如外显子组测序和序列特异性蛋白质组捕获和分析（PICh, HyCCAPP）。然而，通常不可能选择性地洗脱特定捕获的子序列，因为用于双工破坏的条件可能缺乏区分不同序列所需的特异性。我们在这里表明，通过使用支点介导的DNA分支迁移，可以结合和选择性释放多组序列。该策略适用于简单的寡核苷酸混合物，用于交联酵母染色质的序列特异性捕获和特异性释放，以及与DNA微阵列杂交的寡核苷酸的特异性释放。
Nucleic acid hybridization is widely used for the specific capture of complementary sequences from complex samples. It is useful for both analytical methodologies, such as array hybridization (e.g. transcriptome analysis, genetic variation analysis), and preparative strategies such as exome sequencing and sequence-specific proteome capture and analysis (PICh, HyCCAPP). It has not generally been possible to selectively elute particular captured subsequences, however, as the conditions employed for disruption of a duplex can lack the specificity needed to discriminate between different sequences. We show here that it is possible to bind and selectively release multiple sets of sequences by using toehold-mediated DNA branch migration. The strategy is illustrated for simple mixtures of oligonucleotides, for the sequence-specific capture and specific release of crosslinked yeast chromatin, and for the specific release of oligonucleotides hybridized to DNA microarrays.
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