Faculty of Medical Engineering, University Politehnica of Bucharest, Gh Polizu 1-7, 011061 Bucharest, Romania.
Genetic Lab, Str. Milcov, nr. 5, Sector 1, 012273 Bucuresti, Romania.
Molecules. 2023 Jun 7;28(12):4599. doi: 10.3390/molecules28124599.
A nitrocellulose-graphene oxide hybrid that consists of a commercially nitrocellulose (NC) membrane non-covalently modified with graphene oxide (GO) microparticles was successfully prepared for oligonucleotide extraction. The modification of NC membrane was confirmed by Fourier Transform Infrared Spectroscopy (FTIR), which highlighted the principal absorption bands of both the NC membrane at 1641, 1276, and 835 cm (NO) and of GO in the range of 3450 cm (CH-OH). The SEM analysis underlined the well-dispersed and uniform coverage of NC membrane with GO, which displayed thin spider web morphology. The wettability assay indicated that the NC-GO hybrid membrane exhibited slightly lower hydrophilic behavior, with a water contact angle of 26.7°, compared to the 15° contact angle of the NC control membrane. The NC-GO hybrid membranes were used to separate oligonucleotides that had fewer than 50 nucleotides (nt) from complex solutions. The features of the NC-GO hybrid membranes were tested for extraction periods of 30, 45, and 60 min in three different complex solutions, i.e., an aqueous medium, an α-Minimum Essential Medium (αMEM), and an αMEM supplemented with fetal bovine serum (FBS). The oligonucleotides were desorbed from the surface of the NC-GO hybrid membrane using Tris-HCl buffer with a pH of 8.0. Out of the three media utilized, the best results were achieved after 60 min incubation of the NC-GO membranes in αMEM, as evidenced by the highest fluorescence emission of 294 relative fluorescence units (r.f.u.). This value corresponded to the extraction of approximately 330-370 pg (≈7%) of the total oligo-DNA. This method is an efficient and effortless way to purify short oligonucleotides from complex solutions.
一种由商业硝化纤维素(NC)膜非共价修饰氧化石墨烯(GO)微颗粒组成的硝化纤维素-氧化石墨烯杂化体成功地被制备用于寡核苷酸提取。NC 膜的修饰通过傅里叶变换红外光谱(FTIR)得到证实,该光谱突出了 NC 膜在 1641、1276 和 835 cm(NO)以及 GO 在 3450 cm 范围内的主要吸收带(CH-OH)。SEM 分析强调了 NC 膜与 GO 的良好分散和均匀覆盖,其显示出薄的蜘蛛网状形态。润湿性试验表明,NC-GO 杂化膜表现出稍低的亲水行为,水接触角为 26.7°,而 NC 对照膜的接触角为 15°。NC-GO 杂化膜用于从复杂溶液中分离少于 50 个核苷酸(nt)的寡核苷酸。在三种不同的复杂溶液中,即水性介质、α-最小必需培养基(αMEM)和添加胎牛血清(FBS)的αMEM,测试了 NC-GO 杂化膜的提取时间为 30、45 和 60 min 的特性。寡核苷酸从 NC-GO 杂化膜的表面通过 pH 为 8.0 的 Tris-HCl 缓冲液解吸。在所使用的三种介质中,NC-GO 膜在αMEM 中孵育 60 min 后获得了最佳结果,这表现为 294 相对荧光单位(r.f.u.)的最高荧光发射。该值对应于大约 330-370 pg(≈7%)总寡-DNA 的提取。该方法是从复杂溶液中有效且轻松地纯化短寡核苷酸的方法。