Chip-based isothermal amplification devices and methods
AssigneeThe Regents of the University of Michigan
InventorsMark A. Burns, David T. Burke, Brian N. Johnson, John D. DeNuzzio, Wayne F. Beyer, Jr.
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Abstract
Disclosed are methods and compositions for isothermal amplification of nucleic acids in a microfabricated substrate. Methods and compositions for the analysis of isothermally amplified nucleic acids in a microfabricated substrate are disclosed as well. The microfabricated substrates and isothermal amplification and detection methods provided are envisioned for use in various diagnostic methods, particularly those connected with diseases characterized by altered gene sequences or gene expression.
Claims48
A method for isothermal amplification of at least one selected nucleic acid, comprising:
- a) providing at least one sample comprising at least one selected nucleic acid, and reagents effective to permit at least one isothermal amplification reaction, to at least one microfabricated substrate that defines at least one channel, said channel connected to at least one isothermally regulated reaction chamber, wherein said sample is conveyed from said channel to said isothermally regulated reaction chamber by differential heating of said sample, wherein said substrate further comprises at least a second channel or at least one reservoir, wherein said at least a second channel or at least one reservoir is directly or indirectly connected to said at least one reaction chamber, wherein said reagents are prefabricated in a lyophilized form into said at least one reaction chamber, said at least one channel, said at least a second channel or said at least one reservoir;
- and
- b) conducting at least one isothermal amplification reaction to amplify said selected nucleic acid.
A method for isothermal amplification of at least one selected nucleic acid, comprising:
- a) providing at least one sample comprising at least one selected nucleic acid, and reagents effective to permit at least one isothermal amplification reaction, to at least one microfabricated substrate that defines at least one channel, said channel connected to at least one isothermally regulated reaction chamber, wherein said substrate comprises at least one flow-directing means system, wherein said flow-directing means system comprises at least one series of hydrophobic and hydrophilic surface structures arrayed along said channel;
- and
- b) conducting at least one isothermal amplification reaction to amplify said selected nucleic acid.
Dependencies are inferred from the claim text.