Baba Y, Ishimaru N, Samata K, Tsuhako M
Kobe Women's College of Pharmacy, Japan.
J Chromatogr A. 1993 Nov 5;653(2):329-35. doi: 10.1016/0021-9673(93)83191-T.
The performance and the efficiency of several cellulose derivatives as a molecular sieving agent for the capillary electrophoretic separation of DNA restriction fragments were investigated. All fragments up to 12,000 base pairs (bp) in the 1-kbp DNA ladder were resolved using linear polyacrylamide-coated capillaries filled with a buffer solution containing 0.5% cellulose derivative and the separation was completed within 17 min. High-concentration (0.7%) cellulose derivative solutions are effective for the complete separation of small fragments (50-1000 bp) of a HincII and a HaeIII digest of phi X174 DNA. A plate number of 0.5-1 x 10(6) plates per metre was achieved. The migration time and the resolution of DNA fragments were manipulated by varying several parameters, such as the size (viscosity) and the concentration of cellulose derivatives and the applied field strength. Some guidelines are presented for choosing these parameters, depending on the size of the DNA fragments being separated.
研究了几种纤维素衍生物作为毛细管电泳分离DNA限制片段的分子筛剂的性能和效率。使用填充有含0.5%纤维素衍生物的缓冲溶液的线性聚丙烯酰胺涂层毛细管，可分离1-kbp DNA阶梯中所有长达12,000碱基对（bp）的片段，且分离在17分钟内完成。高浓度（0.7%）纤维素衍生物溶液对于完全分离phi X174 DNA的HincII和HaeIII酶切的小片段（50 - 1000 bp）有效。实现了每米0.5 - 1×10⁶ 块板的塔板数。通过改变几个参数，如纤维素衍生物的大小（粘度）、浓度和施加的场强，来控制DNA片段的迁移时间和分辨率。根据待分离DNA片段的大小，给出了选择这些参数的一些指导原则。
