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Transmission of poly(dT60) single‐stranded DNA through polycarbonate track‐etched ultrafiltration membranes

机译:聚(dT60)单链DNA通过聚碳酸酯轨道蚀刻超滤膜的传输

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Abstract The objective of this study was to examine membrane filtration of a single stranded DNA (ssDNA) with 60 thymine nucleotides, and to elucidate the variables controlling its transmission across track‐etched porous membranes. Dead end filtration measurements were performed using different pore size membranes (10, 15, and 30 nm) at different transmembrane pressures in solutions with ionic strength ranging from 0 to 1000 mM NaCl. The diffusivity of the ssDNA was determined using fluorescence recovery after photobleaching, yielding hydrodynamic radii ranging from 1.6 to 2.8 nm, with values decreasing with increasing solution ionic strength. Despite the small ssDNA/membrane pore size, nearly 100 rejection was observed for measurements performed with the 10 and 15 nm pore size membranes under low‐ionic strength conditions. These high rejections can be attributed to strong repulsive electrostatic ssDNA‐membrane interactions. With increasing ionic strength, electrostatic interactions as well as the effective size of the ssDNA decreases and the flexibility of the ssDNA increases, leading to a reduction in ssDNA rejection. A design of experiments approach was used to plan filtration experiments that adequately covered the variable space with a manageable number of experiments. The results yielded an empirical expression relating ssDNA rejection to pore size, solution ionic strength and transmembrane pressure. There was evidence of flow induced elongation at high‐transmembrane pressures in the 30 nm pore size membranes, but not in the smaller pore size membranes. These results are consistent with critical flux estimates developed using a free draining model for the ssDNA.
机译:摘要 本研究的目的是研究具有60个胸腺嘧啶核苷酸的单链DNA(ssDNA)的膜过滤,并阐明控制其在轨道蚀刻多孔膜上传递的变量。在离子强度范围为 0 至 1000 mM NaCl 的溶液中,使用不同孔径的膜(10、15 和 30 nm)在不同的跨膜压力下进行死端过滤测量。使用光漂白后的荧光恢复法测定ssDNA的扩散率,得到1.6至2.8 nm的流体动力学半径,其值随着溶液离子强度的增加而减小。尽管 ssDNA/膜孔径较小,但在低离子强度条件下使用 10 nm 和 15 nm 孔径膜进行的测量观察到接近 100% 的截留率。这些高排斥率可归因于强烈的排斥静电ssDNA-膜相互作用。随着离子强度的增加,静电相互作用以及ssDNA的有效尺寸减小,ssDNA的柔韧性增加,导致ssDNA排斥降低。采用实验设计方法规划过滤实验,以可控的实验数量充分覆盖可变空间。结果产生了将ssDNA截留与孔径、溶液离子强度和跨膜压力相关的经验表达式。有证据表明,在30 nm孔径的膜中,在高跨膜压力动诱导的伸长率,但在较小的孔径膜中没有。这些结果与使用ssDNA的自由排水模型开发的关键通量估计一致。

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