DNA sequencing by capillary electrophoresis using mixtures of polyacrylamide and poly(N,N-dimethylacrylamide).
The possibility of using polymer mixtures with different chemical compositions as a DNA sequencing matrix by capillary electrophoresis (CE) has been exploited. Polyacrylamide (PAM, 2.5%, w/v) having a molecular mass of 2.2 x 10(6) has been mixed with poly(N,N-dimethylacrylamide) (PDMA) having molecular masses of 8000, 470000 and 2.1 x 10(6) at concentrations of 0.2, 0.5 and 1% (w/v). Unlike polymer mixtures of the same polymer with different molecular masses, the use of polymer mixtures with different chemical compositions encounters an incompatibility problem. It was found that the incompatibility increased with increasing PDMA molecular mass and PDMA concentration, which resulted in decreased efficiency in DNA sequencing. Also, the incompatibility had a more pronounced effect on the efficiency as the base number was increased. However, by choosing a low-molecular-mass PDMA of 8000 and a low concentration of 0.2% (w/v), the incompatibility of PAM and PDMA has been alleviated. At the same time, the advantage of using polymer mixtures revealed a higher efficiency for such a polymer mixture when compared with PAM. The mixture also endowed the separation medium with a dynamic coating ability. An efficiency of over 10 x 10(6) theoretical plates per meter has been achieved by using the bare capillaries without the additional chemical coating step.
Song
L
L
Chemistry Department, State University of New York at Stony Brook, 11794-3400, USA.
Liang
D
D
Chen
Z
Z
Fang
D
D
Chu
B
B
D000178:Acrylamides / Q000737:chemistry*; D000180:Acrylic Resins / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D007834:Lasers; D008970:Molecular Weight; D017422:Sequence Analysis, DNA / Q000379:methods*; D013050:Spectrometry, Fluorescence; D000099166:Polyacrylamides
D000178:Acrylamides; D000180:Acrylic Resins; D000099166:Polyacrylamides; C429790:poly(N,N-dimethylacrylamide)