Self-contained biological analysis

A method of amplifying nucleic acids in a sample, comprising the steps of

• a. providing a container having
• i. a plurality of fluidly connected reaction blisters including an amplification blister,
• ii. an additional set of second-stage amplification chambers fluidly connected to the amplification blister, each additional second-stage amplification chamber configured for further amplifying a specific nucleic acid that may be present in the sample, and
• iii. one or more sealable ports fluidly connected to the reaction blisters, the sealable ports providing the only access from an exterior of the container to the reaction blisters such that when all of the one or more sealable ports is sealed, the container is fully closed,
• b. injecting the sample into the container via a first of the sealable ports, and sealing the first port subsequent to injecting the sample,
• c. extracting the nucleic acids from the sample, and moving the extracted nucleic acids to the amplification blister,
• d. subjecting the nucleic acids in the amplification blister to amplification conditions,
• e. fluidly moving a portion of the nucleic acids to each of the additional second-stage amplification chambers,
• f. performing second-stage amplification in the additional second-stage amplification chambers, and
• wherein the extracting step includes lysing cells contained within the sample to generate a lysate,
• mixing nucleic acid-binding magnetic beads with the lysate and incubating to allow the nucleic acids to bind to the magnetic beads,
• deploying a magnet adjacent to one of the plurality of blisters to contain the magnetic beads in that blister, and
• moving the remainder of the lysate to another of the plurality of blisters.