Pipetting

• Pipetting involves drawing a liquid into a pipet and allowing liquid to drain from the pipet in a controlled manner.

• Pipetting is used to quantitatively transfer exact volumes of a liquid from one container to another.

• Mohr pipet

• Has graduations that allow it to deliver a range of volumes.

• Serological pipets have no base mark (the graduations continue onto the tip) and are graduated to deliver.

• That means you let all the (measured) liquid in the pipet exit.

Graduated to the tip

The volumetric pipet has a single graduation that allows it to deliver one specific volume accurately.

Reading a Pipette

Total Volume

Smallest Unit Increment

•Determine the volume of solution in a pipet by reading the bottom of the meniscus at eye level. •Record the volume using all certain digits and one uncertain digit. Certain digits are obtained from calibration marks. •Uncertain digits (the last digit in the number) are estimated between calibration marks.•What are the volumes in these pipets?

          

                                      

          

                                      

                                     

           

Reading the Volume

Reading a Pipet

What is wrong with this pipet reading?

Dispensing Liquid

•Accurately transferring a specified volume of liquid from one container to another requires careful attention to detail. •Pay special attention to the level of liquid in the pipet and carefully monitor the meniscus.

Using a Pipet

3-Way Safety Bulb

Using 3 way bulb

• Make sure there is enough solution in your beaker to completely fill the pipet.

• Never pipet a reagent directly from a reagent bottle.

• Carefully place the attachment of the three-way bulb over the mouth of the pipet.

1. Squeeze the air valve (A) and the bulb simultaneously to empty the bulb of air.

2. Place the tip of the pipet below the solution's surface in the beaker. Gradually squeeze the suction valve (S) to draw liquid into the pipet.

3. If the level of the solution is not high enough, squeeze the air valve (A) and the bulb again to expel the air from the bulb.

4. Draw up more liquid by squeezing the suction valve (S).

5. When the liquid is above the specified volume, stop squeezing the suction valve (S).

6. Do not remove the bulb from the pipet.

7. Do not allow liquid to enter the pipet bulb.

8. Use the expel (E) valve to release the solution.