Method for performing molecular reactions by using immiscible intermediate fluids

Abstract

The present invention relates to a method for performing molecular reactions in a device. The method comprises the steps of (a) introducing one or more reagent solutions and an immiscible intermediate fluid into the device, wherein the device comprises a substrate, on which chemically or biochemically recognizable entities are immobilized; (b) performing molecular reactions between the immobilized chemically or biochemically recognizable entities and the reagent solutions, or on the immobilized chemically or biochemically recognizable entities in the presence of the reagent solutions; (c) displacing the reagent solutions present on the substrate by the immiscible intermediate fluid; (d) separating the immiscible intermediate fluid and the reagent solutions, and reusing the reagent solutions and / or immiscible intermediate fluid for one or more repetitions of steps (a) to (e). The invention further relates to a device for performing the molecular reactions, comprising a reaction zone, reservoirs and liquid connections and a collection and regeneration zone wherein the immiscible intermediate fluid and the reagent solutions are separable by gravitational separation, or a redirection and distribution module, wherein the immiscible intermediate fluid and reagent solutions are separated. The invention also relates to the use of an immiscible intermediate fluid for displacing a reagent solution present in a reaction zone in a microfluidic device, as well as the use of a corresponding device for performing a sequencing reaction or a nucleic acid synthesis reaction.

Description

field of invention [0001] The present invention relates to a method for carrying out molecular reactions in a device comprising the steps of: (a) introducing one or more reagent solutions and an immiscible intermediate fluid into said device, wherein said device comprises a chemical compound immobilized thereon. or a substrate of a biochemically identifiable entity; (b) carrying out a molecular reaction between said immobilized chemically or biochemically identifiable entity and said reagent solution; or in the presence of said reagent solution on said immobilized chemically or biochemically performing a molecular reaction on an identifiable entity; (c) replacing a reagent solution present on said substrate with said immiscible intermediate fluid; (d) separating said immiscible intermediate fluid and said reagent solution; and regenerating One or more repetitions of steps (a) to (e) are performed using said reagent solution and / or immiscible intermediate fluid. The invention ...

AI Technical Summary

Problems solved by technology

One reason for the low sequencing speed is the limited reaction rate when incorporating nucleotides, which is limited by the concentration of enzymes or other reactants in the reaction mixture for cost reasons

What's more, due to the need to wash the entire surface after each step of nucleotide incorporation, all reagents are often wasted

Thus, total reagent consumption is primarily proportional to read length and constitutes a major cost factor

This in turn leads to a tendency to reduce reagent concentrations as much as possible with the disadvantage of longer reaction times

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