Song L, Liang D, Kielescawa J, Liang J, Tjoe E, Fang D, Chu B
Chemistry Department, State University of New York at Stony Brook, 11794-3400, USA.
Electrophoresis. 2001;22(4):729-36. doi: 10.1002/1522-2683(200102)22:4<729::AID-ELPS729>3.0.CO;2-O.
Copolymers of acrylamide (AM) and N,N-dimethylacrylamide (DMA) with AM to DMA molar ratios of 3:1, 2:1 and 1:1 and molecular weights of about 2.2 MDa were synthesized. The polymers were tested as separation media in DNA sequencing analysis by capillary electrophoresis (CE). The dynamic coating ability of polydimethylacrylamide (PDMA) and the hydrophilicity of polyacrylamide (PAM) have been successfully combined in these random copolymers. A separation efficiency of over 10 million theoretical plates per meter has been reached by using the bare capillaries without the additional polymer coating step. Under optimized separation conditions for longer read length DNA sequencing, the separation ability of the copolymers decreased with decreasing AM to DMA molar ratio from 3:1, 2:1 and 1:1. In comparison with PAM, the copolymer with a 3:1 AM:DMA ratio showed a higher separation efficiency. By using a 2.5% w/v copolymer with 3:1 AM:DMA ratio, one base resolution of 0.55 up to 699 bases and 0.30 up to 963 bases have been achieved in about 80 min at ambient temperatures.
合成了丙烯酰胺（AM）与N，N - 二甲基丙烯酰胺（DMA）的共聚物，其中AM与DMA的摩尔比为3:1、2:1和1:1，分子量约为2.2 MDa。这些聚合物在毛细管电泳（CE）的DNA测序分析中作为分离介质进行了测试。在这些无规共聚物中，聚二甲基丙烯酰胺（PDMA）的动态涂层能力和聚丙烯酰胺（PAM）的亲水性已成功结合。使用未经过额外聚合物涂层步骤的裸毛细管，已达到每米超过1000万理论塔板数的分离效率。在针对更长读长DNA测序的优化分离条件下，共聚物的分离能力随着AM与DMA摩尔比从3:1、2:1降至1:1而降低。与PAM相比，AM:DMA比例为3:1的共聚物显示出更高的分离效率。通过使用2.5% w/v的AM:DMA比例为3:1的共聚物，在环境温度下约80分钟内，实现了高达699个碱基时0.55的单碱基分辨率以及高达963个碱基时0.30的单碱基分辨率。
