Automated point-of-care devices for complex sample processing and methods of use thereof

A microfluidic device comprising:

• a reagent-dispensing unit comprising at least one reagent pouch comprising one or more reagents and a sealing layer;
• wherein said at least one reagent pouch comprises at least a first non-compartmentalized reagent pouch comprising an aqueous reagent and a non-aqueous immiscible reagent packaged together in said first non-compartmentalized reagent pouch;
• at least one actuator element comprising spatially oriented magnets, and at least one protrusion configured to apply an actuation force to said at least one reagent pouch to dispense said one or more reagents into a rotatable microfluidic cartridge;
• and
• wherein said rotatable microfluidic cartridge comprises a plurality of wells, is positioned proximate to said at least one actuator element, is configured to receive magnetic beads, and is configured to rotate about a central axis such that in a single actuation step comprising rotating the microfluidic cartridge said spatially oriented magnets capture, re-suspend, and transport said magnetic beads between said plurality of wells.