What Are Matters to Be Paid Attention to When Using Pipette and Its Tips?

1. A complete pipetting cycle
To pipette, there are several steps, installing tip, setting volume, aspirating, dispensing, and removing the tip. The following operating specifications should be followed for each step.

1.1 Pipette tip installation
For single-channel pipettes, insert the tip of the pipette vertically into the tip, press lightly and turn it left and right to tighten; For a multi-channel pipette, align the channel of the pipette with the tip, insert it at an angle, and shake it back and forth slightly to tighten; To prevent it from air leaking, the tip cannot be used repeatedly. If not, it will loosen some parts for the strong impact and what’s worse, it will make the knob of adjusting volume get stuck.

1.2 Capacity setting
When adjusting from a large volume to a small volume, rotate it counterclockwise to the scale; To ensure accuracy, when adjusting from small to the large one, first adjust the set volume clockwise and then return. Adjusting the knob out of the range is forbidden, or the pipette mechanical parts will be damaged.

1.3 Tip aspiration and dispensing
Press the button of the aspiration pipette to the position, release the button, and perform aspiration. Be sure not to move too fast, or the liquid will enter the tip too fast, making the liquid be sucked inside the pipette. Put the liquid close to the wall of the container, first press the first gear, pause briefly and then press the second gear to remove the remaining liquid.

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● Aspirate liquid vertically.
● For 5ml and 10ml pipettes, the sterile filter tips need to be immersed in the liquid surface 5mm, aspirate the liquid slowly, after reaching a predetermined volume, under the liquid pause for 3 seconds, and then leave.
● Slowly release the controller when aspirating liquid, otherwise, the liquid will enter the tip too quickly and the liquid will be sucked into the pipette.
● To absorb volatile liquid, saturate the steam in the sleeve chamber by rinsing the conductive tips 4-6 times in order to avoid liquid leakage.

1.4 Pipette correct placement
Hang the pipette upright on the holder after use. Also, be careful not to fall off. When there is liquid in the pipette tip, do not place the pipette horizontally or upside down, so as to prevent the liquid from flowing back and corroding the piston spring.
If not in use, adjust the range of the pipette tip to a large scale, so that the spring is in a relaxed state to protect the spring.

2. Common wrong operations
1) Hitting the tip repeatedly is forbidden when assembling the tip, as it will make pipetting difficult and even bring damage to it.
2) When aspirating liquid, the pipette tilts, causing inaccurate pipetting, and the liquid easily enters the pipette holder.
3) When aspirating liquid, release your thumb quickly. This will force the liquid to form a turbulent state, and the liquid will directly flush into the pipette.
4) Directly press to the second level.
5) Pipet small volumes of samples with a large range pipette (pipette with appropriate range should be selected).
6) Lay the pipette with residual liquid tips flat (the pipette should be hung on the pipette holder).

3. Notes for pipetting different samples
3.1 Pay attention to two points when removing volatile samples:
a. Be sure to rinse twice before pipetting;
b. Drain the liquid as soon as possible after the suction is completed.

3.2 Pipette high-viscosity samples
Use the reverse pipetting mode: press the suction/discharge button to the second level (second stop point) when aspirating liquid, and press the button to the first level (first stop point) when discharging liquid. In addition, a residence time of 3-5 seconds is required for both suction and discharge.

3. 3 Pipette high-density/low-density samples
The accuracy of the pipette is based on transferring pure water. If the density of the sample is very different from the density of the water, the accuracy will be much worse. Therefore, you need to figure out the density of the sample before pipetting, and then adjust the range to the product of the volume of the sample to be transferred and the density.

For example, if the density of a sample is 1.2g/cm3 and you need to transfer 300ul, then you should set the range to 360ul. This is only a rough adjustment method, and a strict adjustment method requires accurate calculations with the aid of a measuring instrument or a balance as an auxiliary tool.

3. 4 Pipette high temperature/low-temperature samples
Three points need to be paid attention to when removing high temperature/low-temperature samples:
First, never rinse the tip before pipetting;
Second, change a new tip every time you pipette;
Third, liquid aspiration and drainage must be completed as soon as possible.