Integrated purification and measurement of DNA methylation and co-measurement of mutations and/or mRNA expression levels in an automated reaction cartridge

A cartridge for determining a methylation state of a nucleic acid, said cartridge comprising:

• (i) a first column comprising a first matrix material,
• (ii) a sample receiving chamber,
• (iii) a reaction channel or chamber that can be temperature controlled,
• (iv) a plurality of chambers for containing reagents and/or buffers, wherein:
• at least one of said chambers is configured to contain a bisulfite reagent, and
• at least one of said chambers is configured to contain a desulphonation/elution buffer,
• wherein said cartridge optionally comprises a second column comprising a second matrix material,
• wherein said sample receiving chamber, said first and optional second column(s), said plurality of chambers, and said temperature-controlled heating channel or chamber, are selectively in fluid communication by microfluidic channels and valves, and wherein:
• said sample receiving chamber, said first and optional second column(s), said plurality of chambers, and said reaction channel or chamber or a port into said reaction channel or chamber, are disposed around a central valve and selectively in fluid communication with a channel in said central valve, wherein said central valve is configured to accommodate a plunger that is capable of drawing fluid into or out of a chamber in fluid communication with said central valve, and/or
• said cartridge is configured to perform bisulfite conversion in the reaction channel or chamber, which can be subjected to thermocycling and later used for amplification of converted nucleic acid by polymerase chain reaction (PCR).