Journal
ELECTROPHORESIS
Volume 30, Issue 14, Pages 2488-2498Publisher
WILEY
DOI: 10.1002/elps.200800773
Keywords
DNA sequencing; Long read length; Thermal-responsive; Viscosity switchable
Funding
- National Science Foundation, Polymers Program [DMR 0454887]
- National Human Genome Research Institute [NIH 2R01HG01386-09]
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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 similar to 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.
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