Devices and methods for extraction, separation and thermocycling

A method of preparing nucleic acid for analysis using a centrifugal microfluidic device, the method comprising:

• moving a first reagent from a first reagent chamber to an extraction chamber using centrifugal force;
• extracting a first fluid from a sample by heating the sample in the extraction chamber;
• moving the first fluid to a metering chamber using centrifugal force;
• moving the first fluid to a mixing chamber using centrifugal force;
• introducing a second reagent into the mixing chamber through the metering chamber using centrifugal force thereby providing a second fluid;
• moving the second fluid to a reaction chamber using centrifugal force;
• degassing the second fluid using centrifugal force;
• moving the second fluid to a cooling chamber using centrifugal force;
• heating and cooling the second fluid cyclically;
• snap-cooling the second fluid;
• moving a third reagent using centrifugal force to the cooling chamber to form a third fluid;
• loading a polymer into a polymer chamber using centrifugal force;
• and
• mixing the third fluid with the polymer to create a fourth fluid.