Reduced matrix viscosity in DNA sequencing by CE and microchip electrophoresis using a novel thermo-responsive copolymer.
Reduced matrix viscosity in DNA sequencing by CE and microchip electrophoresis using a novel thermo-responsive copolymer.
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DOI：
10.1002/elps.200800773
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发表时间：
2009-07
期刊：
影响因子：
2.9
通讯作者：
Chu, Benjamin
中科院分区：
文献类型：
--
作者：
Wan, Fen;He, Weidong;Zhang, Jun;Chu, Benjamin
高粘度是由亲水性聚合物制成的测序基质的固有问题。这个问题在使用微芯片的分离中被放大，其中通道甚至更小。合成了一种以线型聚丙烯酰胺（LPA）为主链，聚环氧丙烷（PPO）为接枝侧链的新型热缔合接枝共聚物。注入问题可以通过引入PPO侧链来解决，PPO侧链可以通过使用温度变化来组装，但对LPA的筛分能力没有明显的不利影响。粘度测量显示，这些LPA-g-PPO共聚物具有约40 °C的转变温度，在该转变温度以上观察到粘度的显著增加。测序性能取决于PPO的热缔合性质和相关参数。在没有优化的情况下，在ABI 310分析仪上使用1. 8 wt% LPA-g-PPO（1.8MDa，PPO，0.2%）在一小时内实现1000个碱基的读取长度和0.3的单碱基分辨率。这种新型的热可逆共聚物溶液可以作为毛细管电泳中DNA测序的可行基质，在微芯片电泳中更是如此。
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 ~ 40 °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 capillary electrophoresis, and even more so in microchip electrophoresis.
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