As a standardized experimental tool, 96-well plates are widely used in high-throughput experiments and batch operations. Its compact design allows multiple samples to be processed simultaneously, greatly improving experimental efficiency and data processing throughput. However, to complete pipetting operations accurately and quickly in such tiny holes requires not only precision instruments, such as multi-channel pipettes, but also the skilled skills and meticulous operation of the experimenter. Especially when processing 96-well plates, small errors can have a significant impact on experimental results. Therefore, mastering effective 96-well plate pipetting techniques is crucial to ensure the accuracy and reproducibility of experimental data.

1. Preparation work

1. Choose the appropriate pipette:

1>Choose a pipette with appropriate range and accuracy according to experimental needs. For 96-well plates, multi-channel pipettes can significantly improve pipetting efficiency and reduce repeated operations.

2> Make sure the pipette is in good condition and undergo regular calibration and maintenance.

2. Prepare pipetting tips:

1>Choose a pipette tip that matches the pipette to ensure good sealing.

2>Check the suction head for damage or contamination before use and replace it in time.

3. Sample and reagent preparation:

1> Ensure that the quality and purity of samples and reagents meet experimental requirements.

2> The validity period and storage conditions of samples and reagents stored for a long time should be checked.

2. Pipetting operation

1. Install the suction head correctly:

1>Align the pipette tip with a row or column of tips, insert gently and apply pressure evenly to ensure that the tip fits tightly with the pipette.

2>Be careful not to use excessive force to avoid damaging the tip or pipette.

2. Pre-rinse the tip:

1> Before formal pipetting, pre-rinse the tip 2-3 times with the liquid to be pipetted to remove impurities and bubbles inside the tip.

3. Vertical pipetting:

1>Keep the pipette vertical when pipetting to avoid tilting or shaking which may lead to inaccurate pipetting.

2> Gently insert the tip into the liquid, avoiding contact with the bottom or side walls of the container to prevent contamination.

4. Slowly release liquid:

1>When adding liquid to the 96-well plate, gently press the release button of the pipette and slowly release the liquid.

2>Make sure the liquid completely enters the hole to avoid bubbles or splashing.

5. Evenly distribute:

1>For experiments that require precise distribution of liquid, pay attention to controlling the amount of liquid in each hole to be as consistent as possible.

2>Multi-channel pipettes or automatic dispensing systems can be used to ensure uniform distribution.

3. Precautions

1. Avoid cross-contamination:

1> Avoid the tip touching other objects or liquid surfaces during pipetting to prevent cross-contamination.

2>Replace the suction head promptly after use and properly dispose of waste liquid.

2. Temperature control:

1>For experiments that need to be performed at a specific temperature, pay attention to controlling the temperature of the pipetting environment to ensure the accuracy of the experimental results.

3. Cleaning and maintenance:

1>Clean pipettes and tips promptly after use to prevent residues from affecting the next experiment.

2>Calibrate and maintain the pipette regularly to maintain its accuracy and stability.

4. Marking and recording:

1> Mark the position of each well and the corresponding sample or reagent name on the 96-well plate to avoid confusion and errors.

2>Record the experimental process and results in detail for subsequent analysis and summary.

As can be seen from the above, 96-well plate pipetting skills involve many aspects, including preparation, pipetting operations, and precautions. By mastering these skills and strictly abiding by experimental specifications, pipetting accuracy and efficiency can be ensured, thereby improving the reliability and repeatability of experimental results.