Pipette tips, which are considered to be one of the most fundamental laboratory techniques, are utilized in a wide variety of treatments and applications in the field of life science. It is important to note that there are a number of common pipetting errors that can lead to results that are inaccurate and imprecise. The purpose of this all-encompassing guide is to provide a comprehensive overview of ten essential tips and techniques for improving the quality of various experiments and becoming proficient in pipetting. Some of the significant topics that are discussed in this book include temperature equilibrium, pipette angle, pre-wetting, aspiration depth, touch-off techniques, viscosity effects, and volume range optimization. These are just some of the topics that are essential. When pipetting, it is essential to adhere to these best practices in order to enhance the accuracy, precision, and reproducibility of the results.

A Listing of the Items Comprised

It Is Important to Keep the Pipette Angle EvenIn a consistent manner

When attempting to achieve the optimal aspiration depth, touch-off techniques should be utilized.

Liquids that are volatile should be pipetted. It is possible to re-calibrate things in a short amount of time for different densities.

In order to ensure that the pipette, its tips, and the solution have all reached room temperature before beginning the pipetting process, it is essential to wait beforehand. It is possible for the accuracy and precision of the measurement to be negatively impacted by temperature differences between the liquids, the tips, and the equipment. The air cushion that is contained within the pipette can undergo changes in volume as a consequence of the pipette's expansion or contraction in response to changes in temperature. Pipetting should only be done at room temperature if the protocol allows for it, as this will result in the most accurate and reproducible results. The article titled "Effects of Temperature on Pipetting" contains additional information that can be found here.

It is imperative that the pipette angle be maintained consistently.

• There is a possibility that the angle at which the pipette is held can have a significant influence on the accuracy of the pipetting process

• The pipette should be held at a fixed angle that does not exceed twenty degrees from the vertical, as this is the recommended position for the pipette

• When dealing with very small volumes that are measured below 30 microliters, the tilt of the pipette should be kept to a minimum

• Because tilting causes the air cushion pressure inside the tip to change, the aspiration volumes that are produced are not consistent regardless of the angle at which they are produced

• It is recommended that the pipette be held in a vertical position in order to achieve the optimal level of consistency

When you are aspirating, you should submerge the Pipette Tips in the liquid no more than two to three millimeters below the surface of the liquid. As the depth of the immersion increases, there is a greater possibility that the solution will adhere to the surface of the tip. In the event that there is any liquid on the surface of the tip, the volume that is dispensed will be inaccurate. You should take this into consideration. Aspiration of the desired volume can be accomplished in a smooth and even manner by using shallow tip insertion, which is necessary in order to achieve the best possible results.

Making use of techniques that involve touch-off

Immediately following the dispensing process, touch-off techniques should be utilized in order to remove any droplets that are still present on the tip. The following are the three approaches that are suggested for use:1) Side-wall touch off, which involves sliding the tip along the wall of the target vessel; 2) Surface touch-off, which involves touching the droplet to the surface of the liquid; and 3) Direct in-liquid dispensing, which does not require any touch-off beforehand. All three of these methods are described in more detail below. Eliminating any liquid that is still present ensures that the dispense will be completely accurate. Additional information can be found in the article titled "Pipette Touch-Off Techniques."

All the Pre-Wet Advice

Utilize a pipette that falls within the range of the ideal volume.

Always pipette between 35 and 100 percent of the maximum volume of the pipette in order to achieve the highest possible level of accuracy and precision. When the volume of the sample being dispensed is very close to the volume of the pipette as a whole, it is possible to achieve the highest possible level of performance. If the volume of the air cushion is increased below 35%, the degree of variability that results from this increase is increased as well. Maintaining a position within the optimal range is another way to cut down on errors that are dependent on the technique chosen.

When using an electronic pipette to dispense multiple aliquots, it is essential to dispose of the first and last items that are dispensed out of the Pipette Tips. One possible explanation for a low volume is that it is the result of mechanical reversal that occurs after aspiration of the sample. For the final dispense, errors that have been accumulated from earlier dispenses are incorporated into the process. If these two are left out of the equation, it is possible to reduce the variability that exists throughout the dispense series.

Change your approach to liquids that are viscous.

Pipette the viscous samples slowly and at reduced speeds in order to achieve the best results. Allowing for additional pause time is necessary in order to ensure that the liquid is able to enter and exit the tip in its entirety. It is recommended that the reverse pipetting mode be utilized in order to compensate for the adhesion of the inner wall. Any liquid that is still clinging to the tip after the dispensing process is accounted for by the additional volume that is aspirated after the process is complete. The best way to prevent foaming is to avoid blowouts as much as possible.

Pipette substances that are volatile in a short amount of time

When working with volatile liquids like alcohols or aromatics, it is absolutely necessary to make use of pipetting speeds that are relatively fast. It is recommended to use pre-wet tips in order to saturate the air cushion and reduce the amount of evaporation that occurs. Using reverse pipetting allows for the incorporation of additional aspirated volume, which helps to compensate for the volume that is lost due to volatility. Make sure to take the necessary precautions to cut down on the amount of water that drips and evaporates.

Make adjustments in order to take into account the varying densities.

Recalibrating your pipette is something you should do whenever you are working with liquids that have a density that is significantly different from that of water. Calibration of pipettes is typically performed at the factory using water that is at room temperature. This is a standard practice. Inaccurate volume delivery will occur as a consequence of density mismatches if the system that delivers volume is not recalibrated. With regard to pipette calibration, please consult the services.

By putting into practice the ten pipetting tips and best practices that are presented here, you will be able to accomplish your work with an exceptional level of accuracy and precision. You will see an increase in the quality of the results of your experiments if you devote some of your time and energy to perfecting your method.