Wang Qian, Xu Xu, Dai Lixin
Laboratory of Analytical Chemistry, State Key Laboratory of Organometallic Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China.
Electrophoresis. 2006 May;27(9):1749-57. doi: 10.1002/elps.200500597.
The preparation of a new separation matrix, quasi-interpenetrating networks (quasi-IPNs) formed by poly(N-acryloyl-Tris) (poly(tris-A)) and PVP, and its application for dsDNA and ssDNA fragments separation by CE with UV detection, are presented. This new quasi-IPN exhibited high sieving performance, good dynamic coating ability, and low viscosity. Single-base resolutions of dsDNA fragments (Rs = 0.92 for 123/124 bp) and ssDNA fragments (Rs = 0.65 for 123/124 base, Rs = 0.48 for 309/310 base) were achieved by using the quasi-IPN of poly(tris-A)/PVP (2% + 2%) solution in a 31 cm effective length linear polyacrylamide (LPA)-coated column. Single-base separation of dsDNA fragments (Rs = 0.92 for 123/124 bp) was also obtained within 28 min in a 46.7 cm effective length bare column at higher 160 V/cm electric field strength by using the same quasi-IPN solution. The RSD of the migration time measured for each DNA fragments was less than 1.5% in the bare column for nine continuous runs. The effects of temperature and electric field strength on the DNA separation were also investigated.
本文介绍了一种新型分离基质——由聚(N-丙烯酰基三羟甲基氨基甲烷)(聚(三羟甲基氨基甲烷-A))和聚乙烯吡咯烷酮(PVP)形成的准互穿网络(quasi-IPN),及其在紫外检测毛细管电泳中用于双链DNA(dsDNA)和单链DNA(ssDNA)片段分离的应用。这种新型准互穿网络表现出高筛分性能、良好的动态涂层能力和低粘度。通过在31 cm有效长度的线性聚丙烯酰胺(LPA)涂层柱中使用聚(三羟甲基氨基甲烷-A)/PVP(2% + 2%)溶液的准互穿网络,实现了双链DNA片段(123/124 bp时分辨率Rs = 0.92)和单链DNA片段(123/124碱基时Rs = 0.65,309/310碱基时Rs = 0.48)的单碱基分辨率。在46.7 cm有效长度的裸柱中,使用相同的准互穿网络溶液,在160 V/cm的较高电场强度下,28分钟内也实现了双链DNA片段(123/124 bp时Rs = 0.92)的单碱基分离。在裸柱中连续九次运行时,每个DNA片段迁移时间的相对标准偏差(RSD)小于1.5%。还研究了温度和电场强度对DNA分离的影响。