Microfluidics device with continuous flow system, useful e.g. for nucleic acid amplification or sequencing, providing temperature cycling of sample

摘要

A device (A) comprising a microfluidics substrate with at least one pathway for sample flow and at least one heat transfer member (HTM) that can cycle at least part of the sample between at least two temperatures while it flows continuously along the pathway, is new. Independent claims are also included for the following: (1) performing a (bio)chemical process by cycling a flowing sample, containing reagents, between at least two temperatures; (2) performing a biochemical process by injecting at least one reagent, from a reservoir, into a channel carrying a continuous flow of solution containing a sample, and transferring heat between a thermal support and a temperature-regulated part of the channel; (3) performing a temperature cycle on a continuous flow of solution containing a sample; (4) amplifying nucleic acids, comprising: (a) mixing at least one sample comprising the nucleic acids with reagents; (b) feeding at least one channel with a continuous flow of the mixture; (c) running at least one mixture through at least one temperature regulated zone; and (d) cycling the zone through a temperature cycle of at least two temperatures in a predetermined series, so that the nucleic acid undergoes denaturation-hybridization-elongation cycle; and (5) detecting at least one nucleotide in target nucleic acid, comprising: (a) feeding a channel with a continuous flow of a target nucleic acid solution; (b) injecting a reagent for amplifying a region of the nucleic acid which carries the nucleotide to be detected into a channel from a reagent reservoir; (c) running the solution through at least one temperature regulated zone so that the nucleic acid is amplified; (d) injecting the reagent for purifying the amplification product into the channel from a second reagent reservoir; (e) running the solution through at least one temperature regulated zone, to carry out purification; (f) injecting the microsequencing reagent comprising buffer, at least one microsequencing primer, at least on ddNTP and a polymerase into the channel from a third reagent reservoir; (g) running the reaction mixture through at least one temperature regulated zone to produce at least one cycle comprising the denaturation of the target nucleic acid, the hybridization of the nucleic acid with the primer, and the incorporation of the ddNTP which is complementary to the nucleotide to be detected at the 3' end of the primer; and (h) detecting the incorporated ddNTP.