Purpose: The purpose of this study was to examine the pain reducing effects of the dextrose-coated pacifier on venipuncture in premature infants. Methods: The design of this study is a nonequivalent control group pretest-posttest design and a crossover trial. The analysed cases were 40 premature infants (20 in experimental group and 20 in control group) in neonatal intensive care unit of a University Hospital, Gyeongnam Province, Korea. The data were collected from April to October, 2009. The experimental treatment was carried out nursing 20% dextrose-coated pacifier on venipuncture for IV injection. The instruments were O2 saturation and heart rate on pulse oxymeter monitor to measure physiologic pain responses, and NIPS to measure behavioral pain responses. Collected data were analyzed with x2 test, t-test using SPSS program. Results: The effects of the 20% dextrose-coated pacifier were found in the physiologic (only heart rate) and behavioral pain response on venipuncture. Conclusion: These finding is suggested that the dextrose-coated pacifier could be an effective nursing intervention for reducing pain on venipuncture in premature infants.