Oligonucleotide analysis by capillary polymer sieving electrophoresis using acryloylaminoethoxyethanol-coated capillaries.
Capillary polymer sieving electrophoresis using dynamic sieving polymer solutions provides size-based separations of oligonucleotides. The polymer sieving system described here resolves single-stranded oligonucleotides with single-base resolution up to lengths of 30 bases within 10 min. The effect of temperature on the separation of synthetic oligonucleotide standards was examined between 20 degrees C and 40 degrees C, with optimal performance at 35-40 degrees C. By adding urea to the sieving buffer single-base resolution could be extended to 60 bases in about 40 min. Best performance was achieved with capillaries coated with a new monomer, acryloylaminoethoxyethanol. This coating provides the necessary stability to ensure long lifetimes.
Talmadge
K W
KW
Bio-Rad Laboratories, Hercules, CA 94547, USA.
Zhu
M
M
Olech
L
L
Siebert
C
C
D000180:Acrylic Resins / Q000737:chemistry*; D001483:Base Sequence; D019075:Electrophoresis, Capillary / Q000379:methods*; D009838:Oligodeoxyribonucleotides / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D014508:Urea / Q000737:chemistry
D000180:Acrylic Resins; D009838:Oligodeoxyribonucleotides; C089575:poly(N-acryloylaminoethoxyethanol); D014508:Urea