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An accurate method of determining gain coefficients of pulse RF-discharge CO2 laser is developed,which involves the use of both the regular 00°1 and 00°2 laser transitions as probes of CO2 laser. The results indicate that the majority of transitions in discharge have anomalous gain coefficients under RF-discharge condition. This fact has not been generally recognized and the neglect of overlapping transitions can lead to errors in determining rotational temperature.