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DNA sequencing by capillary electrophoresis using copolymers of acrylamide and N,N ‐dimethyl‐acrylamide
Author(s) -
Song Liguo,
Liang Dehai,
Kielescawa Jan,
Liang Jason,
Tjoe Edward,
Fang Dufei,
Chu Benjamin
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200102)22:4<729::aid-elps729>3.0.co;2-o
Subject(s) - copolymer , acrylamide , capillary electrophoresis , polyacrylamide , polymer , electrophoresis , molar ratio , chromatography , materials science , chemistry , polymer chemistry , organic chemistry , catalysis
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.