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===Section II Bisulfite conversion of DNA=== *Add 20ul of sample to 130ul of CT conversion reagent solution in a pcr tube *Vortex the sample to mix *Pulse centrifuge *Perform *98C for 8m *64C for 3.5hr *4C hold *Add 600ul of M binding buffer into a column assembly *Load sample(s) to the column *Close the cap and mix by inverting the column several times *Centrifuge at >10,000g for 30sec *Discard the flow through *Add 100ul of M-Wash Buffer to the column *Centrifuge at full speed for 30sec *Add 200ul of M-Desulphonation buffer to the column and let stand at RT for 20m *Centrifuge for 30sec *Add 200ul of M-Wash buffer to the column and repeat centrifuge *Place the column into a 1.5ml tube *Add 10ul of M-elution buffer directly to the column matrix (Volume can be adjusted, depending on the requirements) *Centrifuge for 30sec to elute the DNA
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