Improve Your Pipetting Accuracy in Just 5 Easy Steps (2024)

FAQs

Improve Your Pipetting Accuracy in Just 5 Easy Steps? ›

The following rules apply to all types of pipettes. Never put a pipette in your mouth. Draw the liquid into the pipette using a rubber bulb or pipette pump. Never withdraw a liquid from a near-empty container.

The following rules apply to all types of pipettes. Never put a pipette in your mouth. Draw the liquid into the pipette using a rubber bulb or pipette pump. Never withdraw a liquid from a near-empty container.

Accuracy (closeness to true value) and precision (consistency of measurements) are vital in scientific experiments. To improve these in the lab, regularly calibrate and maintain equipment, use tools within their appropriate ranges, record significant figures correctly, and take multiple measurements.

Forward and reverse pipetting are two basic pipetting modes that can be used with a mechanical pipette. In forward pipetting, the target volume is aspirated and dispensed, and a separate blowout step is used to completely empty the tip by pressing the plunger to the second stop.

The most common way to check your pipette accuracy is by weighing water. The density of water is 1 g/mL. This means that every microliter (µL) should weigh exactly 0.001 g using a high-precision balance.

Reproducible Experiments: An accurate pipette delivers precise volumes of reagents, samples, or solutions, allowing consistent replication of experiments for reliable scientific findings.

Forward pipetting is generally more accurate, especially when transferring small volumes of liquid, while reverse pipetting is generally more efficient, especially when transferring large volumes of liquid.

How to avoid bubbles when pipetting? ›

Pipetting slowly, using the reverse pipetting technique reduces the risk of bubble formation when transferring viscous solutions (3). Reverse pipetting is also recommended when pipetting small volumes into 96 well micro test plates for ELISA techniques.

Temperature has many effects on pipetting accuracy. The factor that has the greatest effect is the temperature difference between the delivery device and the liquid. The air gap (dead air volume) between the liquid surface and the piston experiences thermal expansion effects unique to the case.

Temperature and pressure affect the volume of the air cushion, which affects pipetting accuracy. Also, volatile solvents can evaporate into the air cushion, which leads to an inaccurate and lower dispensed volume than that displayed on the pipette.

Know proper pipetting techniques: If you're new to pipetting or are a veteran who needs a refresher, make sure you are following these techniques when using a pipette: Practice a smooth and slow motion. Pre-wet the pipette tip. Hold the pipette vertically when drawing in liquid and at a 45-degree angle when dispensing.

Using the same tip(s), aspirate the nominal volume. With the tip(s) immersed 2 mm in liquid, hold the pipette vertically for 30 seconds then, with tip(s) immersed in liquid, dispense all remaining liquid from the tip(s). If liquid level does not drop and no air bubbles are present, continue with validation.