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电渗是CE的基本现象之一,它可以控制组分的迁移速度和方向,进

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Presentation on theme: "电渗是CE的基本现象之一,它可以控制组分的迁移速度和方向,进"— Presentation transcript:

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2 电渗是CE的基本现象之一,它可以控制组分的迁移速度和方向,进

3 如何控制电渗流? 理论: 能影响电渗的因素都有可能用于电渗的控制 实际: 能理想地调节电渗而又不影响分离过程的控制方法目前 不多见.
对于DNA分离及测序来说,pH-8.3,常用管壁涂层(涂覆) 技术.

4 化学涂覆 如聚丙烯酰胺、聚乙烯基吡咯烷酮涂层可在pH9-pH6以内基本消除电渗

5 优点 缺点 抑制熔硅毛细管表面Si-OH解离,修饰表面Zeta电势. 聚合物或硅烷化试剂的取代基覆盖已离解的Si-OH.
增加了内壁表面层附近溶液的粘度,涂层的厚度和密度都将影响到表面   层附近溶液的粘度,从而影响涂层的涂覆效果。 涂层寿命长,稳定性好,分离效率高。 缺点 化学键合的过程中涉及毛细管硅烷化和引发单体聚合两步反应,一方面受两步反应的转化率影响,Si-OH很难被完全屏蔽;另一方面,毛细管内的聚合反应很难控制,容易造成涂层的不均一性、不规整性,甚至阻塞毛细管。 价格昂贵。

6 物理涂覆

7 缺点 优点 涂层制备过程简单易操作,涂覆过程可一步完成,
涂层可再生。这种涂层可以是中性的、带正电荷的、也可以是吸附在带正电的聚合物涂层表面转而带负电荷的。 缺点 物理吸附涂层稳定性不及化学键合涂层.

8 用于DNA分离的聚合物 交联聚合物: 交联聚丙烯酰胺、琼脂糖 非交联聚合物

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11 均聚物            poly-N-hydroxyethylacrylamide(PHEA) 存在的问题:

12 共聚物 a. 无规共聚物 b.嵌段共聚物 理想的筛分介质:高筛分能力、低粘度、自涂覆功能
poly (DEA-co-DMA)  聚 (N, N -二乙基丙烯酰胺- co -N, N - 二甲基丙烯酰胺) poly (AM-co-DMA)  聚(丙烯酰胺- co - N, N - 二甲基丙烯酰胺) poly (AA-co-AG)   聚(丙烯酰胺- co - β-吡喃型葡萄糖) poly(AM-co-AAG)   聚(丙烯酰胺- co -葡糖酰丙胺) poly(NEEA-co-NMEA) 聚(乙氧基乙基丙烯酰胺- co -乙氧基甲基丙烯酰胺)等 b.嵌段共聚物 PEG-(CnF2n+1)2 末端带有碳氟化合物的聚乙二醇 PEO-PPO-PEO 低分子量的聚环氧乙烷和聚环氧丙烷的三嵌段共聚物 BEB/EBE 聚环氧乙烷和聚环氧丁烷的三嵌段共聚物等

13 C. 接枝共聚物 PAM-g-PNIPAM (聚丙烯酰胺-g- 聚N-异丙基丙烯酰胺)
PNIPAM-g-PEO (聚N-异丙基丙烯酰胺-g- 聚环氧乙烷) PAM-g-PDMA (聚丙烯酰胺 -g- 聚N,N- 二甲基丙烯酰胺) PDMA-g-PMMA(聚N,N- 二甲基丙烯酰胺-g-聚甲基丙烯酸甲酯)等 存在的问题:

14 共混聚合物 不同分子量的PEO分离ds 和ss DNA HEC,LPA等不同分子量的混合物也都能成功地对DNA分离或排序 存在的问题:

15 准互穿网络聚合物

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17 Yanmei Wang,Dehai Liang, Jingcheng Hao, Dufei Fang, Benjamin Chu, Electrophoresis 2002, 23,

18 Mechanism of particle formation by inverse microemulsion polymerization of AM/AIBN system. M is monomer acrylamide; I is initiator; R is free radical.

19 Electrophoretic separation of DNA fragments generated on Big Dye TM Terminator Cycle Sequencing Standard with AmpliTaq FS under optimum experimental conditions: 2.5% IPNs, 200 V/cm and room temperature. A-track red, T-track blue, G-track green, and C-track black. Electrophoretic conditions were as follows: effective length 36 cm, total length 41cm, 75-m-i.d., 365-m-o.d. anode buffer 1TTE; cathode buffer 1TTE/7M urea. The sample was injected at a constant electric field of 73 V/cm for 40 s. Yanmei Wang, Dehai Liang, Q. Ying, B. Chu, Electrophoresis 2005, 26,

20 quasi-IPN/GNPs复合介质 initiator LPA GNPs quasi-IPN quasi-IPN/GNPs DMA
Preparation of quasi-IPN/GNPs composite matrices

21 During the preparation of Au colloids, citrate acted as both a reducing agent and a capping agent to avoid aggregation of GNPs. The particle sizes of prepared GNPs could be controlled by the amount of trisodium citrate. TEM images of (A) GNPs20 (~20 nm), (B) GNPs40 (~40 nm), and (C) GNPs60 (~60 nm) in water, and (D) GNPs40 (~40 nm) in quasi-IPN3 polymer solution.

22 DNA sequencing by CE and data analysis
The full four-color (base C, T, A, G) electropherograms of Bigdye Terminator V 3.1 sequencing standard DNA sample by CE using 2.5% w/v quasi-IPN3/GNPs40-1 at 50℃. Sequencing conditions: effective/total length of bare fused-silica capillaries, 50/61 cm; id/od, 75/365μm; sequencing electric field strength, 150 V/cm; DNA electrokinetic injection, 41 V/cm for 30 s; anode buffer, 1×TTE; cathode buffer, 1×TTE/7M urea.

23 The partial four-color (base C, T, A, G) electropherograms of standard DNA sample using 2.5% w/v quasi-IPN3/GNPs40-1 by CE at 50℃. Electrophoresis, 2007, 28, Electrophoresis, 2007, 28, 中国发明专利:申请号: 公开号:CN A

24 quasi-IPN/functionalized multi-walled carbon nanotubes
Schematic representation of preparation of MWNT-COOH, MWNT-Br and MWNT-PDMA via ATRP Schematic representation of the formation of quasi-IPN/MWNT-PDMA double-network composite sieving matrix. Resolution vs. base number in DNA sequencing by CE using quasi-IPN Electrophoresis, 2008, 29, TEM images of (A) crude MWNTs in water and (B) MWNT-PDMA in quasi-IPN solution.

25 Formation of HEC-g-PAM
Viscosity versus temperature for different HEC-g-PAM copolymers. .Resolution versus size of DNA fragments by using HEC, HEC-g-PAM and PDMA as separation medium at 1.5% w/v. Schematic representation of the structure of HEC-g-PAM copolymer and of its adsorption to a capillary wall. Electrophoresis, 2007, 28,

26 价格表 毛细管 (61cm/根) 100 cm/根 POP6 3.5 mL 分离介质 5mL ¥1000 ¥ 64 ¥ 2376 ¥ 100


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