Multiphasic DNA adsorption to silica surfaces under varying buffer, pH, and ionic strength conditions.

Multiphasic DNA adsorption to silica surfaces under varying buffer, pH, and ionic strength conditions.
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在不同的缓冲液,pH和离子强度条件下,多最多DNA吸附到二氧化硅表面。

DOI:
10.1021/jp3017776
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发表时间:
2012-05-17
影响因子:
3.3
通讯作者:
Niemz, Angelika
Niemz, Angelika
中科院分区:
化学3区
文献类型:
--
作者:
Vandeventer, Peter E.;Lin, Jessica S.;Zwang, Theodore J.;Nadim, Ali;Johal, Malkiat S.;Niemz, Angelika

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DNA 和硅胶之间的可逆相互作用用于从复杂样品中固相萃取和纯化 DNA。离液盐通常会促进 DNA 与二氧化硅结合,但会抑制 DNA 聚合酶扩增。我们通过大量消耗和石英晶体微天平 (QCM) 实验,在有或没有离液盐的条件下研究了 DNA 对二氧化硅的吸附。虽然使用离液盐将更多的 DNA 吸附到硅胶上,但在某些不含离液盐的缓冲条件下,也产生了相似量的洗脱 DNA。 QCM 结果表明,在较强的吸附条件下,吸附的 DNA 层最初是刚性的,但在几分钟内变得粘弹性。这些结果在质量上与多相吸附过程的数学模型一致。不需要离液盐的缓冲条件可以简化核酸样品制备的方案。了解 DNA 如何吸附到硅胶上有助于优化临床诊断和研究应用的核酸样品制备。
Reversible interactions between DNA and silica are utilized in the solid phase extraction and purification of DNA from complex samples. Chaotropic salts commonly drive DNA binding to silica, but inhibit DNA polymerase amplification. We studied DNA adsorption to silica using conditions with or without chaotropic salts through bulk depletion and Quartz Crystal Microbalance (QCM) experiments. While more DNA adsorbed to silica using chaotropic salts, certain buffer conditions without chaotropic salts yielded a similar amount of eluted DNA. QCM results indicate that under stronger adsorbing conditions, the adsorbed DNA layer is initially rigid, but becomes viscoelastic within minutes. These results qualitatively agreed with a mathematical model for a multiphasic adsorption process. Buffer conditions that do not require chaotropic salts can simplify protocols for nucleic acid sample preparation. Understanding how DNA adsorbs to silica can help optimize nucleic acid sample preparation for clinical diagnostic and research applications.
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