Reversible and Irreversible Mechanical Damaging of Large Double-Stranded DNA upon Electrospraying.
Reversible and Irreversible Mechanical Damaging of Large Double-Stranded DNA upon Electrospraying.
复制标题
电喷雾对大双链 DNA 的可逆和不可逆机械损伤。
DOI：
10.1021/acs.analchem.6b01642
复制
发表时间：
2016
影响因子：
7.4
通讯作者：
V. Morozov
中科院分区：
文献类型：
--
作者：
Y. M. Shlyapnikov;E. Shlyapnikova;V. Morozov
DNA溶液的电流体动力喷雾（或电喷雾，ES）是用于质谱、微阵列制造和DNA纳米气溶胶的产生的有吸引力的技术。在这里，我们报告ES如何影响DNA结构，并评估可能的方法来减少ES对DNA的损伤。结果表明，在任何ES条件下，线性λ-噬菌体DNA受到强烈的断裂，产生片段的混合物。除了这样的片段，显着的可逆性变化的DNA结构中揭示了DNA电泳迁移率略有增加。片段化的程度被证明是减少与减少DNA长度和增加通过ES毛细管的流速。短于5 kbp的片段在ES时没有显示任何显著的损伤。ES过程中作用于DNA的力的实验数据和理论估计都表明DNA被机械力损坏，并且损坏发生在泰勒锥尖端附近，可能是由于在那里操作的高剪切应力或/和粘性阻力。λ-DNA与六氨合钴（III）离子的缩合完全保护了它免受ES的任何损伤。
Electrohydrodynamic spraying (or electrospaying, ES) of DNA solutions is an attractive technique for applications in mass spectrometry, in microarray fabrication, and in generation of DNA nanoaerosols. Here we report how ES affects DNA structure and evaluate possible ways to reduce DNA damage upon ES. It is shown that under any ES conditions, linear λ-phage DNA is subjected to intensive rupture producing a mixture of fragments. In addition to such fragmentation, notable reversible changes in the DNA structure were revealed by a slight increase in DNA electrophoretic mobility. The degree of fragmentation was shown to decrease with decreased DNA length and with increased flow rate through the ES capillary. Fragments shorter than 5 kbp did not show any notable damage upon ES. Both experimental data and theoretical estimations of the forces acting on DNA during ES indicate that DNA is damaged by mechanical forces, and the damage takes place in the vicinity of the Taylor cone tip, presumably due to the high shear stress or/and viscous drag forces operating there. Condensation of λ-DNA with hexamminecobalt(III) ions completely protected it from any damage upon ES.
影响因子：
16.6
作者：
Bullerjahn, Jakob T.;Sturm, Sebastian;Kroy, Klaus
通讯作者：
Kroy, Klaus
影响因子：
4.7
作者：
Xie J;Jiang J;Davoodi P;Srinivasan MP;Wang CH
通讯作者：
Wang CH