Repeated pipetting method is to accurately control the volume of liquid by repeating the pipetting operation multiple times. This method is particularly suitable for handling liquids that are volatile, expensive, or require strict volume control. By repeatedly pipetting, scientists can gradually approximate the required liquid volume, thereby minimizing errors and improving the accuracy and reliability of experiments.
1. Repeat the pipetting operation steps 1. Set the volume: Fully press the pipette button to the second stop point to set the required pipetting volume. 2. Aspirate liquid: Install the matching tip on the pipette and make sure the connection is tight and leak-free. Dip the tip evenly into the liquid to be aspirated, just enough to avoid bubbles. Slowly slide the control button back into place, allowing the liquid to be drawn into the tip. After releasing the button, the tip should be filled with the required volume of liquid. 3. Discharge liquid: Move the suction head above the receiving container, and ensure that the suction head is close to the wall of the container to reduce liquid splashing. Press the drain button to the first stop, where most of the liquid will be drained. Pay attention to controlling the discharge speed to avoid liquid splashing. 4. Repeat pipetting: According to the experimental needs, repeat steps 2 and 3 to perform multiple pipetting operations. Before each pipetting, ensure that the pipette has been reset to the required volume and check that the tips are clean and undamaged. 5. Dispose of remaining liquid: After all pipetting operations are completed, a small amount of liquid usually remains in the pipette. According to the experimental needs, this part of the remaining liquid can be discarded (directly release the button to let the remaining liquid flow out and discarded) or returned to the original storage container (but ensure that the operation will not contaminate the original solution). 6. End the operation: After processing the remaining liquid, release the discharge button to the first stop point and release the button. At this time, the pipette has returned to the initial state. Carefully remove the tips from the receiving container and dispose of according to laboratory regulations. Finally, perform necessary cleaning and disinfection of the pipette for next use.
2. Precautions When performing repeated pipetting, special attention should be paid to controlling the discharge of liquid and the disposal of remaining liquid to ensure the accuracy of pipetting. Clean and calibrate your pipettes regularly to maintain stable performance. When working with liquids of different properties, it may be necessary to change to a different type of tip or adjust pipette settings.
3. Advantages of repeated pipetting method 1. Effective handling of volatile liquids: For volatile liquids, a single pipetting may cause the volume to decrease due to rapid evaporation of the liquid, thus affecting the accuracy of the experiment. By repeatedly pipetting, only a small portion of the liquid can be processed at a time, reducing the time the liquid is exposed to air, thus reducing evaporation losses. In addition, repeated pipetting can also gradually stabilize the liquid during multiple operations, reducing errors caused by volatilization. 2. Reduce liquid loss and contamination: During repeated pipetting, since each operation is relatively independent, liquid loss can be more easily monitored and controlled. By precisely controlling the volume and number of pipettes each time, liquid waste can be minimized. At the same time, since the chance of contact between the liquid and the external environment (such as air, dust, etc.) is reduced, the risk of the liquid being contaminated is also reduced. 3. Precisely control the amount of liquid: Repeated pipetting allows the experimenter to adjust the volume of liquid multiple times according to experimental needs, thereby controlling the amount of liquid more accurately. This precise control is especially important for experiments that require strict volume control (such as drug formulation, chemical reactions, etc.). Through multiple pipetting operations, the required liquid volume can be gradually approached and the accuracy of the experiment can be improved.
4. Disadvantages of the repeated pipetting method 1. Cumbersome operation: The repeated pipetting operation is relatively cumbersome and requires repeating the same steps multiple times, which increases the workload and time cost of the experiment. This tediousness may be more obvious when performing a large number of pipetting operations, affecting the efficiency of the experiment. 2. Increased risk of contamination: During repeated pipetting, the number of contacts between the tip and pipette and the liquid increases, which may increase the risk of contamination. If the tips or pipettes are not cleaned and disinfected promptly and thoroughly, contaminants may be introduced into the experimental system, affecting the accuracy of the experimental results. 3. High skill requirements for operators: Repeated pipetting operations require operators to have a high skill level and stability to ensure the accuracy and consistency of each pipetting. If the operator's skills are insufficient or the operation is improper, pipetting errors may increase and even affect the reliability of experimental results.
With its unique multiple operation mode, the repeated pipetting method provides scientists with an efficient means to effectively control liquid volume. In actual operation, we mastered every step of setting the measuring range, aspirating liquid, discharging liquid, and handling remaining liquid. These steps are crucial and require us to be meticulous in our operations. By repeatedly pipetting, we can not only effectively reduce the evaporation loss of liquid and reduce the risk of contamination, but also ensure the accuracy and reliability of the experiment, thereby laying a solid foundation for the success of scientific research.
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