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First, the tube is filled with the culture solution, then a certain amount of the culture solution is absorbed with a medical syringe, the air in it is drained, the needle is extended to the bottom of the tubule (to prevent the formation of bubbles), and the culture solution is slowly pushed into the tubule until it bulges (the effect of surface tension), and then it is so excessive that the culture solution wetting the tubule surface. Because of the constant viscosity of the culture medium, it can make the wet Duchenne tube attached to the test tube put people slowly slide to the bottom of the test tube, without breaking the bottom of the test tube. After sterilization, it can be used for the actual determination of coliform. After many tests and repeated comparisons, it has been proved that this method can improve the accuracy of experimental observation results and improve the work efficiency.