Microfluidic methods for non-thermal nucleic acid manipulations
AssigneeCaliper Technologies Corp.
InventorsMichael Knapp, John Wallace Parce, Luc J. Bousse, Anne R. Kopf-Sill
Life of this patent
Estimating the term…
Abstract
Integrated systems, apparatus, software, and methods for performing biochemical analysis, including DNA sequencing, genomic screening, purification of nucleic acids and other biological components and drug screening are provided. Microfluidic devices, systems and methods for using these devices and systems for performing a wide variety of fluid operations are provided. The devices and systems of are used in performing fluid operations which require a large number of iterative, successive or parallel fluid manipulations, in a microscale, or sealed and readily automated format.
Claims34
A method of amplifying a target nucleic acid by a non-thermal polymerase chain reaction, the method comprising:
- providing a microfluidic device which includes a reaction chamber or channel, one or more sources of a target nucleic acid and primer sequences, a source of a chemical denaturant and a source of polymerase enzyme fluidly connected to the reaction chamber or channel, and a fluid direction system for delivering the chemical denaturant or the polymerase enzyme to the reaction chamber or channel;
- flowing the target nucleic acid and the primer sequences along the reaction chamber or channel;
- melting complementary strands of the flowing target nucleic acid by delivering a volume of the chemical denaturant to the reaction chamber or channel;
- annealing the primer sequences to the flowing target nucleic acid by eliminating a denaturing effect of the chemical denaturant;
- extending the primer sequences along the flowing target nucleic acid by delivering a volume of the polymerase enzyme to the reaction chamber or channel;
- and
- repeating the steps of melting, annealing and extending to amplify the flowing target nucleic acid.
A method of producing an amplified nucleic acid in a microfluidic device, the method comprising:
- providing a microfluidic device comprising a microscale channel;
- flowing a double stranded nucleic acid in the microscale channel;
- flowing a denaturant into the microscale channel and contacting the flowing double stranded nucleic acid, thereby providing a flowing denatured nucleic acid;
- removing the denaturant;
- and
- amplifying the flowing denatured nucleic acid to produce an amplified nucleic acid.
Dependencies are inferred from the claim text.