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Pipetting Techniques
Best pipetting practice

1) Correct pipette/tip combination 2) Correct immersion depth 3) Vertical pipette posture for liquid uptake
Reduce errors by choosing a pipette/tip combination > Immerse as little as possible The hydrostratic pressure changes with the holding
with the smallest possible air cushion. → preventing liquid transfer on the tip’s outside angle of the pipette.
> Immerse deeply enough to avoid uptake of air

Small air cushion reduces errors: False pipetting when holding 100–1,000 µL Pipette
Setting 100 µL → optimal pipette 10–100 µL Volume in μL Depth in mm at an > 45° angle (set volume = 1,000 µL)
1,008
Measurement error [%]

3.00 0.1–1 1
1,006

Volume [µL]
1–100 2–3 1,004
2.00

100–1,000 2–4 1,002

1.00
1,000
1,000–10,000 3–6
0.00 998
10–100 20–200 100–1,000 0 1 2 3 4 5 6 7 8 9 10
Pipette model [µL] Pipetting steps
systematic error random error Correct pipetting posture: vertical False pipetting posture: > 45°

4) Prewetting = Saturating of air cushion 5) Correct liquid discharge 6) Slow and continuous working pace
Prewet tip at least three times to equilibrate > Discharge liquid by touching the wall of the target To ensure precise and accurate pipetting results
air cushion to liquid. vessel → adhesion force of liquid
> Volumes below 10 µL: discharge directly into the liquid

Pipette set to 100 µL: The first and

the second pipetting step is insufficient
100.4

100.0
Volume [µL]

99.6

99.2

98.8

98.4
1 2 3 4 5 6 7 8 9 10
Pipetting steps

Forward and reverse pipetting

Transfer Forward pipetting Reverse pipetting

1. Press operating button down to 1st stop 1. Press operating button down to 2nd stop
Liquid uptake
2. Let operating button move up completely 2. Let operating button move up completely

Liquid discharge 3. Press operating button via 1st stop down to 2nd stop 3. Press operating button down to 1st stop

Observation 4. No liquid is left in tip after action 4. Liquid is left in tip after action (volume of blow-out)

1. 2. 3. 4. 1. 2. 3. 4.
REST POSITION REST POSITION

1st STOP 1st STOP

2nd STOP 2nd STOP

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