APPARATUS AND METHODS FOR INTEGRATED SAMPLE PREPARATION, REACTION AND DETECTION

Abstract - 2014224115 A method comprising: (a) providing a composition comprising nucleic acids bound to magnetic particles in a first chamber of a cassette comprising the first chamber and a reaction chamber; (b) eluting the nucleic acids from the particles in the first chamber to obtain an eluted nucleic acid sample; (c) magnetically attracting the magnetic particles to a cavity of a valve member movably disposed within a valve housing, wherein the valve housing is at least partially disposed between the first chamber and a second chamber and comprises a first opening in fluid communication with the first chamber and a second opening in fluid communication with the second chamber; (d) transferring at least a portion of the eluted nucleic acid sample to the reaction chamber in fluid communication with the first chamber; and (e) amplifying one or more target nucleic acid sequences in the eluted nucleic acid sample by combining the eluted nucleic acid sample and PCR reagents in the reaction chamber under conditions suitable for PCR. In a preferred embodiment, the PCR master mix contains PCR reagents and substantially no MgCl2 is disposed in the reaction chamber, and the nucleic acids are eluted in an elution buffer comprising MgCl2.