Separation of oligonucleotides and DNA fragments by capillary electrophoresis in dynamically and permanently coated capillaries, using a copolymer of acrylamide and β-D-glucopyranoside as a new low viscosity matrix with high sieving capacity

被引:22
作者
Chiari, M
Damin, F
Melis, A
Consonni, R
机构
[1] CNR, Inst Biocalysis & Mol Recognit, I-20131 Milan, Italy
[2] CNR, Ist Chim Macromol, NMR Lab, I-20133 Milan, Italy
关键词
acrylamide; beta-D-glucopyranoside copolymer; DNA oligonucleotide separations; polydimethylacrylamide; dynamic coating;
D O I
10.1002/elps.1150191817
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
New copolymers of acrylamide and beta-D-glucopyranoside were synthesized and characterized. The different reactivity of the two monomers towards radical polymerization meant we could control the growth of the polymer chains whose length was inversely related to the number of glucose residues incorporated in the copolymers. The properties of these polymers were investigated in the separation of oligonucleotides and double-stranded DNA by capillary electrophoresis (CE) in coated and uncoated capillaries. The new copolymers were a suitable matrix for CE due to their high-resolving capacity and low viscosity. We also looked into the advantages of a new method of dynamic suppression of electroosmotic flow based on the addition of small amounts (0.03-0.05%) of dimethylacrylamide to the sieving and to the running buffer. A complete test was run on the reproducibility and efficiency of separations carried out in a permanently and dynamically coated capillary, and the advantages and disadvantages of the two methods were compared.
引用
收藏
页码:3154 / 3159
页数:6
相关论文
共 31 条
[1]   Analysis of disease-causing genes and DNA-based drugs by capillary electrophoresis - Towards DNA diagnosis and gene therapy for human diseases [J].
Baba, Y .
JOURNAL OF CHROMATOGRAPHY B, 1996, 687 (02) :271-302
[2]   HIGH-RESOLUTION SEPARATION OF DNA RESTRICTION FRAGMENTS BY CAPILLARY ELECTROPHORESIS IN CELLULOSE DERIVATIVE SOLUTIONS [J].
BABA, Y ;
ISHIMARU, N ;
SAMATA, K ;
TSUHAKO, M .
JOURNAL OF CHROMATOGRAPHY A, 1993, 653 (02) :329-335
[3]   THE USE OF COATED AND UNCOATED CAPILLARIES FOR THE ELECTROPHORETIC SEPARATION OF DNA IN DILUTE POLYMER-SOLUTIONS [J].
BARRON, AE ;
SUNADA, WM ;
BLANCH, HW .
ELECTROPHORESIS, 1995, 16 (01) :64-74
[4]   CAPILLARY ELECTROPHORESIS OF DNA IN UNCROSS-LINKED POLYMER-SOLUTIONS [J].
BARRON, AE ;
SOANE, DS ;
BLANCH, HW .
JOURNAL OF CHROMATOGRAPHY A, 1993, 652 (01) :3-16
[5]   Analysis of modified oligonucleotides by capillary electrophoresis in a polyvinylpyrrolidone matrix coupled with electrospray mass spectrometry [J].
Barry, JP ;
Muth, J ;
Law, SJ ;
Karger, BL ;
Vouros, P .
JOURNAL OF CHROMATOGRAPHY A, 1996, 732 (01) :159-166
[6]  
BUTLER JM, 1994, J CHROMATOGR A, V658, P71
[7]  
BUTLER JM, 1994, BIOTECHNIQUES, V17, P4
[8]   Rapid DNA sequencing of more than 1000 bases per run by capillary electrophoresis using replaceable linear polyacrylamide solutions [J].
Carrilho, E ;
RuizMartinez, MC ;
Berka, J ;
Smirnov, I ;
Goetzinger, W ;
Miller, AW ;
Brady, D ;
Karger, BL .
ANALYTICAL CHEMISTRY, 1996, 68 (19) :3305-3313
[9]   POLYMERASE CHAIN-REACTION HETERODUPLEX POLYMORPHISM ANALYSIS BY ENTANGLED SOLUTION CAPILLARY ELECTROPHORESIS [J].
CHENG, J ;
KASUGA, T ;
MITCHELSON, KR ;
LIGHTLY, ERT ;
WATSON, ND ;
MARTIN, WJ ;
ATKINSON, D .
JOURNAL OF CHROMATOGRAPHY A, 1994, 677 (01) :169-177
[10]   Synthesis and characterization of capillary columns coated with glycoside bearing polymer [J].
Chiari, M ;
DellOrto, N ;
Gelain, A .
ANALYTICAL CHEMISTRY, 1996, 68 (17) :2731-2736