A micro-fluid reaction vessel includes an upper plate formed of an elastomer, a lower plate adhered to the upper plate, a micro-chamber and a micro-channel formed on an inner surface of the upper plate facing the lower plate and an inlet hole and an outlet hole formed in the upper plate and through which a fluid flows into or out of, respectively. The micro-channel is constructed to be closed by pressure applied to the upper plate and elastically restored when the pressure is not applied. A micro fluid reaction method uses the micro fluid reaction vessel and a method of manufacturing forms the microfluid reaction vessel.