摘要
合成了一种水溶性聚合物 ,聚N ,N 二甲基丙烯酰胺 (PDMA)和聚乙烯吡咯烷酮体系 (PVP) ,并用于DNA分离 .优选该体系溶液含量为 2 0 %PDMA + 2 0 %PVP ,在紫外检测的条件下 ,对标准DNA酶切片段pBR3 2 2 /HaeIII样品进行分离 ,其中 12 3 / 12 4碱基对的分离度 (Rs)为 0 72 .加入甲酰胺 (φ≥ 0 10 )对DNA样品进行去活处理后 ,该体系亦能对单链DNA片段中的 12 3 / 12 4进行分离 ,分离度 (Rs)为 0 75 .实验还显示进样时存在浓缩效应 .本体系粘度较小 ,筛分能力强 ,具备芯片毛细管DNA分离介质的潜力 .
The water-soluble mixtures of polymers, poly(N,N-dimethylacrylamide) (PDMA)/polyvinylpyrrolidone (PVP), were synthesized and used in capillary electrophoresis (CE) for DNA separation. With the optimal concentration of PDMA/PVP polymer solution, the doublet of 123/124 bp in pBR322/HaeIII Markers could be separated with UV detector and the resolution value was 0.72. When the denaturant formamide (φ≥0.10) was added in the polymer solution, DNA fragments of 123/124 could be also separated well with the resolution as 0.75, which quite met the requirement of DNA sequencing (R s>0.59). In addition, the lower viscosity of the polymer solution made the practice easier. The sample stacking effect was validated in the experiments. The medium showed single-base-pair DNA separation by capillary electrophoresis even with ultraviolet detector and uncoated capillary column.
出处
《化学学报》
SCIE
CAS
CSCD
北大核心
2004年第1期66-70,共5页
Acta Chimica Sinica
基金
中国科学院创新工程重大项目基金 (No.KSCX1 0 6)资助
关键词
单碱基分离
DNA分离
毛细管电泳
紫外检测器
无胶筛分
合成水溶性聚合物
capillary electrophoresis, non-gel sieving, synthetic polymer, DNA separation, UV detector, single-base-pair separation
