Sequence-based separation of single-stranded DNA using nucleotides in capillary electrophoresis: focus on phosphate.
Sequence-based separation of single-stranded DNA using nucleotides in capillary electrophoresis: focus on phosphate.
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DOI：
10.1002/elps.201200683
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发表时间：
2013-06
期刊：
影响因子：
2.9
通讯作者：
McGown, Linda B.
中科院分区：
文献类型：
--
作者：
Zhang, Xueru;McGown, Linda B.
DNA分析在生物学、医学、生物技术和法医学等领域有着广泛的应用。DNA的长度分离是很容易实现使用筛选凝胶电泳。按序列分离就不那么简单了，通常需要在测量之前杂交的链的天然或诱导构象或热或化学稳定性方面有足够的差异。我们之前在运行缓冲液中使用鸟苷-5 ' -单磷酸（GMP）证明了四个单链DNA 76-mers的分离，这些DNA 76-mers仅基于序列的几个a - g取代。我们将分离归因于GMP独特的自组装以形成高阶结构。在这里，我们研究了一组扩展的76-mers，旨在探索分离的机制和实验条件的影响。我们惊奇地发现，其他核糖核苷酸也实现了与GMP相似的分离，并且使用磷酸钠代替GMP实现了一些分离。磷酸钾的分离效果几乎和核糖核苷酸一样好。这表明分离介质为DNA提供了一个物理化学环境，以序列选择的方式影响链迁移。需要进一步的研究来确定这种机制是否涉及磷酸盐和DNA链之间的特定相互作用，或者是分离介质的其他特性的结果。在毛细管凝胶电泳DNA分离中，磷酸盐通常被避免使用，因为它的高离子强度会加剧焦耳加热。我们的结果表明，磷酸化合物应该检查DNA的分离基于序列。
DNA analysis has widespread applicability in biology, medicine, biotechnology and forensics. DNA separation by length is readily achieved using sieving gels in electrophoresis. Separation by sequence is less simple, generally requiring adequate differences in native or induced conformation or differences in thermal or chemical stability of the strands that are hybridized prior to measurement. We previously demonstrated separation of four single-stranded DNA 76-mers that differ by only a few A-G substitutions based solely on sequence using guanosine-5′-monophosphate (GMP) in the running buffer. We attributed separation to the unique self-assembly of GMP to form higher order structures. Here we examine an expanded set of 76-mers designed to probe the mechanism of the separation and effects of experimental conditions. We were surprised to find that other ribonucleotides achieved the similar separation to GMP, and that some separation was achieved using sodium phosphate instead of GMP. Potassium phosphate achieved almost as good separations as the ribonucleotides. This suggests that the separation medium provides a physicochemical environment for the DNA that effects strand migration in a sequence-selective manner. Further investigation is needed to determine whether the mechanism involves specific interactions between the phosphates and the DNA strands or is a result of other properties of the separation medium. Phosphate generally has been avoided in DNA separations by capillary gel electrophoresis because its high ionic strength exacerbates Joule heating. Our results suggest that phosphate compounds should be examined for separation of DNA based on sequence.
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影响因子：
64.5
作者：
FISCHER, SG;LERMAN, LS
通讯作者：
LERMAN, LS
DOI：
10.1073/pnas.86.8.2766
发表时间：
1989-04-01
影响因子：
11.1
作者：
ORITA, M;IWAHANA, H;SEKIYA, T
通讯作者：
SEKIYA, T
影响因子：
1.8
作者：
Spindler, L;Drevensek-Olenik, I;Mariani, P
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影响因子：
7
作者：
Tian, HJ;Brody, LC;Landers, JP
通讯作者：
Landers, JP
影响因子：
2.9
作者：
Chang, Chun Yaw (Joel);Stellwagen, Nancy C.
通讯作者：
Stellwagen, Nancy C.