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Review Your Pipette Technique for Accuracy. Accurate pipetting is a skill that lab technicians should always review and ensure it's done correctly. When pipetting small volumes, ensure you maintain the correct vertical positions and pause for at least one second after aspirating the sample. Pull the pipette straight out from the container, hold ...
If you’re pipetting 2µL, use a 2.5µL pipette instead of a 10µL one. Also, make sure that the tips you’re using fit snugly and reliably, and consider using thin tips for small wells, such as those found in 384-well plates. 2. Check that your pipettes are calibrated. Be sure the pipettes you’re using are also calibrated appropriately.
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Sep 19, 2018 · The following points are essential: 1. Pipette and tip size: Always choose the pipette with the lowest nominal volume possible and the smallest tip to keep the air cushion as small as possible. When pipetting 1 µL e.g., choose a 0.25 – 2.5 µL pipette and matching tip rather than a 1 – 10 µL pipette. 2.
Choosing the correct pipetting technique helps securing the accuracy and precision necessary for reproducible, reliable results. Especially when pipetting small volumes the influence of the pipetting technique can have tremendous effects on the experimental result. Two main techniques exist and each should be applied depending on the sample liquid.
- Choose The Right Pipette and Pipette Tips
- Check That Your Pipettes Are Calibrated
- Dilute Highly Concentrated Stock Solutions
- Avoid Foaming and Bubbles
- Thoroughly Mix The Contents of The Well
- Prevent Evaporation by Covering Your Plate
Volumetric errors are amplified at such low volumes. To mitigate these errors, be sure to select the proper pipette that delivers volumes that match your protocol. As a rule of thumb, select the smallest pipette capable of handling the volume you need. Pipetting accuracy decreases the further your pipetted volume is to the maximum volume of the pip...
Be sure the pipettes you’re using are also calibrated appropriately. You can do a quick check using distilled water and a microbalance. Since the density of water is 1g/mL, each microliter of distilled water at room temperature should weigh 1mg.
Working with highly concentrated stock solutions requires working with even lower volumes. If your wells can handle additional volume, prepare intermediate solutions. For example, work with 2X or 5X stock solutions instead of 10X if you can.
Whether you’re weighing the benefits of popping bubbles with the pipette tip (without introducing more bubbles!), or you’re resigned to starting over, there are some preemptive measures you can take to avoid this problem in the first place. Try reverse pipetting. Although this requires a little extra volume, bubbles and foaming are greatly reduced....
When handling small volumes, mixing is particularly important as handling mistakes are much more obvious. When mixing by pipetting, be sure to set the volume on your pipette equal to at least half the volume in the well you want to mix. While this can be done manually, you can save a lot of finger fatigue by learning how to set mixing programs on a...
Evaporation is always an issue when handling liquids, and the effects are magnified when working with small volumes. Use plastic plate covers after you are done or if you need to take a break. The plate remains covered until you are ready to use it. Alternatively, you can use parafilm or sealing tape to help prevent evaporation. Although pipetting ...
May 7, 2017 · Keep in mind that analytical balances are far more accurate than any pipette. Dispense volumes appropriate for your instrument, but at the same time use the largest volume possible for your experiment. For example, consider diluting samples to deliver the same quantity of reagent in a larger volume. Good practice vs. best practice.
Before aspirating, it is important that you immerse the tip properly below the meniscus. For large volume pipettes (1-5 mL), the immersion depth should be at least 5-6 mm, while for smaller volume pipettes, the depth should be around 2-3 mm. If you immerse it too little, specifically the large volume pipettes, it can result in the aspiration of ...
