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Are transit systems required to take paratransit reservations during normal business hours?

Yes. Under Department of Transportation (DOT) Americans with Disabilities Act (ADA) regulations at 49 C.F.R. Section 37.131(b), paratransit reservation service must be available during at least all normal business hours of the entity’s administrative offices, as well as during times, comparable to normal business hours, on a day when the entity's offices are not open before a service day (e.g., on a Sunday).