The following are some key points to remember when pipetting, they will help to improve your technique and results:
It is important to ensure you do not immerse the pipette tip to far below the surface as it can cause the air in the tip to become compressed and too much liquid to be drawn up.
The pipette should be held vertically whilst pipetting, once again, to ensure too much liquid is not drawn up, which would result in inaccurate measurements.
When dispensing, water tension can prevent some of the remaining liquid in the tip from exiting, in order to avoid this happing, the tip of the pipette should be held against the inside of the vessel to draw out the remaining liquid.
The pipette tip should be pre-rinsed 2 or more times to give it a clean and to coat the inside to make the volume dispensed more accurate.
The plunger should be compressed and released slowly to ensure all the air is displaced when compressing and no air bubbles are drawn up when releasing.
Liquids should be pipetted at room temperature as temperature can affect the volume of the liquid. Pipette, tip and liquid should all be at the same temperature to increase accuracy.
The two main techniques for pipetting are:
- Is a great technique to use for viscous liquids, liquids with a tendency to foam and for very small volumes.
- It is performed by pushing the plunger down to the second stop, putting the tip into the liquid and releasing the plunger to the original position. To dispense the liquid, push the plunger down to the first stop and then either disposing of the tip or dispensing the remainder of the liquid into the waste, or its original container by the pushing the plunger to the second stop again.
- Is a great technique for mixing a reagent or sample into another liquid and recommended for aqueous solutions.
- It is performed by pushing the plunger down to the first stop, putting the tip into the liquid and releasing the plunger back to the original position. To dispense the liquid, push the plunger down to the first stop, then after a short pause press down to the second stop to empty the tip. The tip can then be disposed of or used again.
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