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，进而影响 CE 的分离效率和重现性．

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 .

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

优点•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。

物理涂覆

• 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。

优点

缺点物理吸附涂层稳定性不及化学键合涂层．

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

均聚物

　　　　　　　　　　　 poly-N-hydroxyethylacrylamide （ PHEA)

存在的问题：

共聚物

a. 无规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

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：

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：

准互穿网络聚合物

Yanmei Wang,Dehai Liang, Jingcheng Hao, Dufei Fang, Benjamin Chu, Electrophoresis 2002, 23, 1460-1466

M

M

R

M

M

M

M

M M

MM

M

M

R

M

R

M

M

M

M

M

MM

M

M

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

18 19 20 21 22 230

2x106

4x106

250

20015010050CT T CT C CT CA CTG T CT C CACTTCCTTG ACAAGT GC CGC T C ATG CT T C TTTTG C CT C CCG CTG CT C CGA A AGA CTAGG C CG CGA AT CAG A A C CA C CACAG T CA ATA T CA C C A C CTT C CT CTT AT AG AT T CGG AA T CT CATG AT AG G G G CT CAG C C T C

24 25 26 27 28 290

2x106

4x106

400

350300TG TG CG AG TG G AG AG AAG TT T G CAG G CG AG CTG AGG AG CA A T TG CAG G TG A T A TG ATG TG C T CGG C T CA AG AAG CG GG C C CG G AG AGG AAG AAG T CG TG C CG G G G C T A A T T A T TG G C A A A ACG AG C T C T TG T TG T A A A C A T T G A T C C A A C TG G A A TG T

30 31 32 33 34 350

2x106 550500450

A C T A A TG G C G A A T CA A T A T T C CA T A AG G CA TG A TG G T TG C T CAG AGG C AGG AG AAG AG C AA CG A A TA C G A T C C T A T A A A AG A T A A A A CAT A A A T A A A CAG T C T TG A TT A T A T T CTG G G T A T T A A AG C C A C A A T CAG A A C A A A T A T A TG CT T TG T AT C T

36 37 38 39 40 41 420

1x106

2x106

750

700650600

AG TAC AG AG AC A G A G G G C G T T AG T A A T G T G AA T G T A C GT T TAGT G T A G T AG AG A T A C A A A A ACTG A G ACGTATG GA A T A T T ACT A CT A T G T A CTA CGG A T A CA T CAGTTA ATGATGAG TG A G AGG T

T T CT TG C C T T CT T CA T T A C CA A CTG C T T C CG CGG CC A C A TT A AG AG A C TTG TGG T A A G ATAA G A G ATA TTTT A TT CG TT CTG CTG A C T G C T GG ATG T C GG AAA TA T CTG C A TT G AT AAG AG CG T A A T G CAG A T AT A T G TAG TG A A A G T CG TG C TAT G T A C G G A G

43 44 45 46 47 480

1x106

2x106

850800

migration time, min

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 1TTE; cathode buffer 1TTE/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, 126-136

quasi-IPN

GNPs

quasi-IPN/GNPs

DMA initiator

quasi-IPN LPA

Preparation of quasi-IPN/GNPs composite matrices

quasi-IPN/GNPs复合介质

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.

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.

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 qu

asi-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.

DNA sequencing by CE and data analysis

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, 1072-1080. Electrophoresis, 2007, 28, 2998-3007

中国发明专利：申请号： 200610114099.3. 公开号： CN1966719A

1) HNO3

COOH

COOH

2) SOCl23) HOCH2CH2OH

Br Br

O4)

COOCH2CH2OOCC BrCH3

CH3

COOCH2CH2OOCC BrCH3

CH3

COOCH2CH2OOCCCH3

CH3

CH2 CH Brm

C=ON

H3C CH3

COOCH2CH2OOCCCH3

CH3

CH2 CH Brm

C=ON

H3C CH3

DMA

CuBrMe6[14]aneN4

MWNT MWNT-COOH MWNT-Br MWNT-PDMA

Electrophoresis, 2008, 29, 4637-4645.

+

MWNT-PDMA

quasi-IPN quasi-IPN/MWNT-PDMA double-network matrix

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.

TEM images of (A) crude MWNTs in water and (B) MWNT-PDMA in quasi-IPN solution.

Resolution vs. base number in DNA sequencing by CE using quasi-IPN

OH

Hydroxyethyl cellulose Hydroxyethyl cellulose Hydroxyethyl cellulose

OHOH OHO O

BrO

O O

NH2

BrO

O O

NH2

OHO O

Br Br

O

Br

Br acrylamide

ligand,cat.

OH

Hydroxyethyl cellulose Hydroxyethyl cellulose Hydroxyethyl cellulose

OHOH OHO O

BrO

O O

NH2

BrO

O O

NH2

OHO O

Br Br

O

Br

Br

nn

TEA, DMAP

■ HEC○ HEC-g-PAM-1▲ HEC-g-PAM-2

◇ HEC-g-PAM-3▼ HEC-g-PAM-4

Electrophoresis, 2007, 28, 3223-3231.

HEC-g-PAM

Formation of HEC-g-PAM

Viscosity versus temperature for different HEC-g-PAM copolymers.

Schematic representation of the structure of HEC-g-PAM copolymer and of its adsorption to a capillary wall.

.Resolution versus size of DNA fragments by using HEC, HEC-g-PAM and PDMA as separation medium at 1.5% w/v.

价格表

毛细管(61cm/ 根 )

毛细管100 cm/ 根

POP6

3.5 mL

分离介质5mL

¥1000 ¥ 64 ¥ 2376 ¥ 100