Table 1.

Heating and Cooling Times of the Two Temperature-Sensitive Steps in the GlycoWorks Protocol (Denaturation and De-N-Glycosylation) during the Manual and Automated Protocols.

	Manual			Automated
	Heating Time (min)	Cooling Time (min)	Total Time (min)	Heating Time (min)	Cooling Time (min)	Total Time (min)	Heating Time Difference (Automated – Manual)	Cooling Time Difference (Automated – Manual)	Total Time Difference (min)
Denaturation at 90 °C	3	3	6	16	15	31	13	12	25
De-N-glycosylation at 50 °C	5	3	8	3	5	8	−2	2	0

During the manual protocol, the user has two heating blocks preset to the desired temperatures, allowing them to drop the samples in and take them out at the allotted times. The automated protocol, however, requires that the samples stay in a Peltier effect heating block at room temperature throughout the reactions. This block then heats to the desired temperature and cools back down to room temperature.