Sequential Injection Analysis for Optimization of Molecular Biology Reactions

Abstract

In order to automate the optimization of complex biochemical and molecular biology reactions, we developed a Sequential Injection Analysis (SIA) device and combined this with a Design of Experiment (DOE) algorithm. This combination of hardware and software automatically explores the parameter space of the reaction and provides continuous feedback for optimizing reaction conditions. As an example, we optimized the endonuclease digest of a fluorogenic substrate, and showed that the optimized reaction conditions also applied to the digest of the substrate outside of the device, and to the digest of a plasmid. The sequential technique quickly arrived at optimized reaction conditions with less reagent use than a batch process (such as a fluid handling robot exploring multiple reaction conditions in parallel) would have. The device and method should now be amenable to much more complex molecular biology reactions whose variable spaces are correspondingly larger.

Autonomous microfluidic actuators for periodic sequential flow generation

“…Control of sequential periodic flows in microfluidic chips has numerous applications, such as layer-by-layer assembly (1, 2), sequential-injection analysis (3,4), soft robot actuation (5)(6)(7), and spatiotemporal analysis of cells (8)(9)(10)(11)(12) and organisms (13)(14)(15). These applications, however, have been mainly implemented by dynamic off-chip controllers that require many external connections and instructions for users.…”