Department of Polymer Science and Engineering, CAS Key Laboratory of Soft Matter Chemistry, University of Science and Technology of China, Hefei 230026, China.
Talanta. 2009 Nov 15;80(1):195-201. doi: 10.1016/j.talanta.2009.06.048. Epub 2009 Jul 1.
A new matrix additive, poly (N,N-dimethylacrylamide)-functionalized gold nanoparticle (GNP-PDMA), was prepared by "grafting-to" approach, and then incorporated into quasi-interpenetrating network (quasi-IPN) composed of linear polyacrylamide (LPA, 3.3 MDa) and PDMA to form novel polymer/metal composite sieving matrix (quasi-IPN/GNP-PDMA) for DNA sequencing by capillary electrophoresis. Without complete optimization, quasi-IPN/GNP-PDMA yielded a readlength of 801 bases at 98% accuracy in about 64 min by using the ABI 310 Genetic Analyzer at 50 degrees C and 150 V/cm. Compared with previous quasi-IPN/GNPs, quasi-IPN/GNP-PDMA can further improve DNA sequencing performances. This is because the presence of GNP-PDMA can improve the compatibility of GNPs with the whole sequencing system, enhance the entanglement degree of networks, and increase the GNP concentration in system, which consequently lead to higher restriction and stability, higher apparent molecular weight (MW), and smaller pore size of the total sieving networks. Furthermore, the composite matrix was also compared with quasi-IPN containing higher-MW LPA and commercial POP-6. The results indicate that the composite matrix is a promising one for DNA sequencing to achieve full automation due to the separation provided with high resolution, speediness, excellent reproducibility, and easy loading in the presence of GNP-PDMA.
一种新的基质添加剂,聚(N,N-二甲基丙烯酰胺)-功能化的金纳米粒子(GNP-PDMA),通过“接枝到”方法制备,然后掺入到由线性聚丙烯酰胺(LPA,3.3 MDa)和 PDMA 组成的准互穿网络(quasi-IPN)中,形成用于毛细管电泳 DNA 测序的新型聚合物/金属复合筛分基质(quasi-IPN/GNP-PDMA)。在不完全优化的情况下,quasi-IPN/GNP-PDMA 在 50°C 和 150V/cm 下使用 ABI 310 遗传分析仪约 64 分钟即可达到 98%的准确率,读取长度为 801 个碱基。与以前的 quasi-IPN/GNPs 相比,quasi-IPN/GNP-PDMA 可以进一步提高 DNA 测序性能。这是因为 GNP-PDMA 的存在可以改善 GNPs 与整个测序系统的兼容性,增强网络的缠结程度,并增加系统中的 GNP 浓度,从而导致更高的限制和稳定性、更高的表观分子量(MW)和更小的总筛分网络的孔径。此外,还将复合基质与含有更高分子量 LPA 的 quasi-IPN 和商业 POP-6 进行了比较。结果表明,由于在存在 GNP-PDMA 的情况下提供了高分辨率、快速、出色的重现性和易于加载的分离,该复合基质是用于 DNA 测序实现全自动化的有前途的基质之一。