Microfabricated bioprocessor for integrated nanoliter-scale Sanger DNA sequencing
Microfabricated bioprocessor for integrated nanoliter-scale Sanger DNA sequencing
复制标题
DOI：
10.1073/pnas.0602476103
复制
发表时间：
2006-05-09
影响因子：
11.1
通讯作者：
Mathies, Richard A.
中科院分区：
文献类型：
--
作者：
Blazej, Robert G.;Kumaresan, Palani;Mathies, Richard A.
一种高效的纳米级微加工生物处理器已经被开发和评估，该处理器集成了所有三个Sar ger测序步骤：热循环、样品纯化和毛细管电泳法。混合玻璃-聚二甲基硅氧烷(PDMS)晶片规模结构用于将250-NL反应器、亲和力捕获净化室、高性能毛细管电泳道以及气动阀门和泵整合到单个微型制造设备上。单芯片实验室级别的集成使仅用1 fmol的DNA模板即可完成Sanger测序。对多达556个连续碱基进行了测序，准确率为99%，证明了人类和其他完整(K)基因组从头测序所需的阅读长度。这种小型化DNA测序仪的性能为预测桑格测序的最终成本和效率极限提供了基准。
An efficient, nanc liter-scale microfabricated bioprocessor integrating all three Sar ger sequencing steps, thermal cycling, sample purification, and capillary electrophoresis, has been developed and evaluated. Hybric glass-polydimethylsiloxane (PDMS) wafer-scale construction is used to combine 250-nl reactors, affinity-capture purification chambers, high-performance capillary electrophoresis channels, and pneumatic valves and pumps onto a single microfabricated device. Lab-on-a-chip-level integration enables complete Sanger sequencing from only 1 fmol of DNA template. Up to 556 continuous bases were sequenced with 99% accuracy, demonstrating read lengths required for de novo sequencing of human and other complete (K) genomes. The performance of this miniaturized DNA sequencer provides a benchmark for predicting the ultimate cost and efficiency limits of Sanger sequencing.