通过微加工技术合成的用于 DNA 吸附的核碱基修饰微凝胶

Kemal Çetin
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摘要

DNA 分离是一个关键步骤,因为基于 DNA 的检测在分子生物学、生物化学和生物医学应用中具有重要意义。本研究的目的是制造微米大小的水凝胶作为 DNA 的吸附剂。在 N,N'-亚甲基双丙烯酰胺作为交联剂的存在下,通过自由基聚合法在微模板阵列芯片的微孔中合成了聚(2-羟乙基甲基丙烯酸酯-甲基丙烯酸缩水甘油酯)微凝胶。然后,通过甲基丙烯酸缩水甘油酯的环氧基团将腺嘌呤固定在微凝胶上。扫描电子显微镜和傅立叶变换红外光谱被用来研究微凝胶的化学和形态特征。通过吸附研究确定了核酸酶固定化微凝胶吸附 DNA 的最佳条件。DNA吸附在微凝胶上的数量在最初增加后,在DNA浓度约为2.0毫克/毫升时达到饱和水平。在最佳 pH 值和温度条件下,初始 DNA 浓度为 2.0 毫克/毫升时,微凝胶的最大吸附量为 38.54 毫克/克。在反复的吸附-解吸循环中,DNA 吸附能力没有明显下降。研究结果表明,腺嘌呤固定化微凝胶是吸附 DNA 的可行选择。
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Nucleobase-modified microgels synthesized via microfabrication technology for DNA adsorption
DNA isolation is a crucial procedure since DNA-based assays have great importance in molecular biology, biochemistry and biomedical applications. The objective of this study is to fabricate micron-sized hydrogels as adsorbents for DNA. Poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) microgels were synthesized by free radical polymerization in the presence of N,N'-methylenebisacrylamide as a crosslinker, in the microholes of a microstencil array chip. Then, adenine was immobilized to microgels through the epoxy groups of glycidyl methacrylate. Scanning electron microscopy and Fourier transform infrared spectroscopy were employed to investigate the chemical and morphological characterizations of the microgels. Adsorption studies were carried out to determine the optimal conditions for DNA adsorption of nucleobase-immobilized microgels. After initially increasing, the quantity of DNA adsorbed onto the microgels reached a saturation level at a DNA concentration of around 2.0 mg/mL. The maximum adsorption was 38.54 mg/g microgels for an initial DNA concentration of 2.0 mg/mL in the optimum medium pH and temperature. DNA adsorption capabilities are shown to not significantly decline in recurrent adsorption-desorption cycles. As a result of the findings, adenine-immobilized microgels were demonstrated to be a viable option for DNA adsorption.
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