Sequential DEXAS: a method for obtaining DNA sequences from genomic DNA and blood in one reaction.
Sequential DEXAS: a method for obtaining DNA sequences from genomic DNA and blood in one reaction.
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顺序 DEXAS：一种在一次反应中从基因组 DNA 和血液中获取 DNA 序列的方法。
DOI：
10.1093/nar/gng112
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发表时间：
2003
影响因子：
14.9
通讯作者：
C. Kilger
中科院分区：
文献类型：
--
作者：
M. Motz;G. Sagner;S. Pääbo;C. Kilger
描述了连续 DEXAS（直接指数扩增和测序），这是一种一步扩增和测序程序，可以直接从基因组 DNA 中准确、廉价且快速地确定 DNA 序列。该方法依赖于同时使用两种 DNA 聚合酶，这两种酶在掺入双脱氧核苷酸的能力以及在反应过程中被激活的时间上都不同。一种酶掺入脱氧核苷酸并进行靶 DNA 序列的扩增，以活性状态提供，而另一种酶掺入双脱氧核苷酸并进行测序反应，则以非活性状态提供，但在热循环过程中通过温度步骤而被激活。因此，在反应的初始阶段，发生靶标扩增，而在第二阶段发生测序反应。我们证明，Sequential DEXAS 可以直接从基因组 DNA 以及直接从人类血液中产生高质量的测序结果，无需事先分离或纯化 DNA。
Sequential DEXAS (direct exponential amplification and sequencing), a one step amplification and sequencing procedure that allows accurate, inexpensive and rapid DNA sequence determination directly from genomic DNA, is described. This method relies on the simultaneous use of two DNA polymerases that differ both in their ability to incorporate dideoxynucleotides and in the time at which they are activated during the reaction. One enzyme, which incorporates deoxynucleotides and performs amplification of the target DNA sequence, is supplied in an active state whereas the other enzyme, which incorporates dideoxynucleotides and performs the sequencing reaction, is supplied in an inactive state but becomes activated by a temperature step during the thermocycling. Thus, in the initial stage of the reaction, target amplification occurs, while in the second stage the sequencing reaction takes place. We show that Sequential DEXAS yields high quality sequencing results directly from genomic DNA as well as directly from human blood without any prior isolation or purification of DNA.
基因组 DNA 的耦合扩增和测序。
DOI：
10.1073/pnas.88.7.2815
发表时间：
1991
影响因子：
11.1
作者：
Ruano,G;Kidd,KK
通讯作者：
Kidd,KK
影响因子：
56.9
作者：
Fliss, MS;Usadel, H;Sidransky, D
通讯作者：
Sidransky, D