Reduced matrix viscosity in DNA sequencing by CE and microchip electrophoresis using a novel thermo-responsive copolymer.
High viscosity is an inherent problem of sequencing matrices made of hydrophilic polymers. This problem is amplified in separations using microchips where the channels are even smaller. A novel thermal-associating graft copolymer, using linear polyacrylamide (LPA) as the backbone and poly(propylene oxide) (PPO) as the graft side chain was synthesized. The injection problem could be resolved by introducing PPO side chains that can be assembled by using temperature changes but without an obvious detrimental effect on the sieving ability of the LPA. Viscosity measurement showed that these LPA-g-PPO copolymers had a transition temperature of approximately 40 degrees C, above which a significant increase in viscosity was observed. The sequencing performance depended on the thermal association properties of PPO and related parameters. Without optimization, a read length of 1000 bases with a single base resolution of 0.3 was achieved within an hour on an ABI 310 analyzer, using 1.8 wt% LPA-g-PPO (1.8 MDa, PPO, 0.2%). This novel thermal reversible copolymer solution can be a promising candidate as a viable matrix for DNA sequencing in CE, and even more so in microchip electrophoresis.
Wan
Fen
F
Department of Chemistry, Stony Brook University, Stony Brook, NY, USA.
He
Weidong
W
Zhang
Jun
J
Chu
Benjamin
B
D000178:Acrylamides / Q000737:chemistry; D000180:Acrylic Resins / Q000737:chemistry*; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D019075:Electrophoresis, Capillary / Q000379:methods*; D046209:Electrophoresis, Microchip / Q000379:methods*; D008027:Light; D011108:Polymers / Q000737:chemistry*; D011409:Propylene Glycols / Q000737:chemistry*; D012542:Scattering, Radiation; D017422:Sequence Analysis, DNA / Q000379:methods*; D013696:Temperature; D014783:Viscosity; D000099166:Polyacrylamides
D000178:Acrylamides; D000180:Acrylic Resins; D004247:DNA; D011108:Polymers; D011409:Propylene Glycols; D000099166:Polyacrylamides; C052970:poly-N-isopropylacrylamide; C012504:polypropylene glycol