Electro-hydrodynamic extraction of DNA from mixtures of DNA and bovine serum albumin
Electro-hydrodynamic extraction of DNA from mixtures of DNA and bovine serum albumin
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从 DNA 和牛血清白蛋白混合物中电流体动力学提取 DNA
DOI：
10.1039/d0an00961j
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Ladd, Anthony J.
中科院分区：
文献类型：
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作者：
Valley, Benjamin E.;Crowell, Anne D.;Butler, Jason E.;Ladd, Anthony J.
我们报道了通过压力驱动流和平行电场之间的电-流体动力耦合从牛血清白蛋白（BSA）中分离基因组DNA （48 kbp）。电流体动力提取利用这种耦合在微流体收缩通道的入口捕获DNA分子，同时允许蛋白质和盐从设备中冲洗出来。将含有λ-DNA （1 ng）和BSA （0.3 mg）的样品（10 μL）直接注射到装置中，并通过流动的缓冲溶液对流到收缩通道入口。当蛋白质和盐被洗脱时，DNA基本上无限期地被困在这个区域。该概念的有效性已通过DNA和牛血清白蛋白浓度的荧光测量进行了评估。在单级装置中发现电流体动力萃取可将DNA浓度提高40倍，同时将BSA浓度降低4个数量级。收缩通道入口处DNA与BSA的相对浓度可达1.5:1，A260/280比值为1.9。无盐溶液中DNA的最大产率为50%，而盐溶液（150mm）的产率较低（38%）。
We report separation of genomic DNA (48 kbp) from bovine serum albumin (BSA) by the electro-hydrodynamic coupling between a pressure-driven flow and a parallel electric field. Electro-hydrodynamic extraction exploits this coupling to trap DNA molecules at the entrance of a microfluidic contraction channel, while allowing proteins and salts to be flushed from the device. Samples (10 μL) containing λ-DNA (1 ng) and BSA (0.3 mg) were injected directly into the device and convected to the contraction channel entrance by a flowing buffer solution. The DNA remains trapped in this region essentially indefinitely, while proteins and salts are eluted. The effectiveness of the concept has been assessed by fluorescence measurements of DNA and BSA concentrations. Electro-hydrodynamic extraction in a single-stage device was found to enhance the concentration of DNA 40-fold, while reducing the BSA concentration by four orders of magnitude. The relative concentrations of DNA to BSA at the contraction channel entrance can be as large as 1.5 : 1, corresponding to an A260/280 ratio of 1.9. The maximum yield of DNA from a salt-free solution is 50%, while salted (150 mM) solutions have a lower yield (38%).