Microfluidic methods for manipulating dna

Abstract

Techniques are provided for generating, manipulating, and measuring fluidic droplets in mixed phase systems based on establishing transient continuities between otherwise spatially separated phases. In certain methods of the invention, electrodes in contact with the continuous phases allow electrical monitoring of continuity or proximity of separated phases as a means to characterize droplets. In other methods of the invention, fluidic continuity provides a means for generating droplets, injecting or extracting the contents of droplets, and sorting droplets. Chemical techniques are also provided that use these droplet-based methods, or others, to quantify and identify nucleic acids through incorporation into hydrogel particles. The nucleic acids are entrapped either actively by chemical incorporation during gel polymerization or passively by chain entanglement. After incorporation into the hydrogel particles, the nucleic acids are solvent accessible either at the particle periphery or within internal pores for further biochemical manipulations and characterization. In one important aspect, the invention combines the high specificity, high sensitivity, and unbiased performance of clonal DNA amplification in free solution with the simplicity of permanently co-localizing the separate reaction products onto rigid substrates for myriad biochemical applications.