Automated bead-trapping apparatus and control system for single-molecule DNA sequencing

被引：6

作者：

Bashford, Greg ^{[1
]}

Lamb, Don ^{[2
]}

Grone, Dan ^{[2
]}

Eckles, Bob ^{[2
]}

Kornelsen, Kevin ^{[3
]}

Middendorf, Lyle ^{[2
]}

Williams, John ^{[2
]}

机构：

[1] Univ Nebraska, Dept Biol Syst Engn, Lincoln, NE 68583 USA

[2] LI COR Biosci Inc, Lincoln, NE 68504 USA

[3] Micralyne Inc, Edmonton, AB T6N 1E6, Canada

来源：

OPTICS EXPRESS | 2008年 / 16卷 / 05期

关键词：

D O I：

10.1364/OE.16.003445

中图分类号：

O43 [光学];

学科分类号：

070207 ; 0803 ;

摘要：

We have been investigating a microfluidics platform for highspeed, low-cost sequencing of single DNA molecules using novel "charge-switch" nucleotides. A significant challenge is the design of a flowcell suitable for manipulating bead-DNA complexes and sorting labeled polyphosphate molecules by charge. The flowcell is part of a single-molecule detection instrument, creating fluorescence images from labeled polyphosphates. These images would ultimately be analyzed by signal processing algorithms to identify specific nucleotides in a DNA sequence. Here we describe requirements of the fluidics system for loading, identifying, tracking, and positioning beads. By dynamically modulating pressure gradients in the plenum chambers of a multi-channel network, we could guide individual beads with high precision to any desired coordinate and reversibly trap them in stepped channels. We show that DNA immobilized on pressure-trapped beads can be physically extended into a downstream channel under electric force for analysis. Custom dynamic algorithms for automated bead control are described. (C) 2008 Optical Society of America.

引用

页码：3445 / 3455

页数：11

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