Micropipetting Practice

1) List the steps of placing 20 uL into a solution in order. Please refer to the demo.

You would start by getting comfortable with the 2 stops when pushing down the top. Once comfortable, you would then
open the box of tips, and push down the micropipette onto the tip. The box must be closed immediately, as to keep the
environment as sterile as possible. You must then secure the tip, then you can begin with the liquid. Push down to the
first stop then place the tip in at an angle in the middle (not touching any sides) of the solution. From there, push down
to the second stop and release the top slowly in order to avoid air bubbles. Bring the pipette to the second solution and
stick the tip in the middle of the solution. Push down to both stops and do not release until the tip is completely out of
the solution. Then release and take off the tip for proper disposal.

There are different sizes of micropipettes. Choosing the right size is a MUST. On the side or top of each micropipette
there is a range that is appropriate for that amount.

In the example to the left, you’ll see that there is a 20-200 uL range. This means
the smallest volume it can contain is 20 uL and the largest is 200 uL.

The general rule of thumb is to choose the pipette that best fits your range,
without it going too close to the highest number. For example, if I wanted to
transfer 200uL of solution, I would use a 200-500uL pipette rather than the one
shown to the left.

2) Now that you have chosen the correct micropipette, it is time to set the proper amount
of liquid within the appropriate range to draw up/transfer. This is done with a dial on the
side that is either vertical or horizontal (school’s ones are vertical). They look like the
images below.

To read and set the dials assume the uppermost/leftmost number is the first
place of the LARGEST amount the micropipette can hold. So let’s look at the
leftmost pipette. At the top you can see that it’s range is 100-1000uL. 1000uL is
the largest amount it can hold and so the topmost box would stand for the
thousands place. This means that if the numbers were set from top to bottom as
1-0-0-0, it would draw up 1000 uL.

How much liquid then is

each of the images
drawing up based on
the numbers in the boxes?

a) 550

b) 35

c) 1.2
 3) Now it’s time for you to practice. When you are working with MICRO solutions (total solution volume being the
size of a teardrop) then even a minute error can result in a BIG difference. Draw the amounts in the boxes on the
micropipette and the type of micropipette you would use on the line below each one.

Micropipette Options: 1-5 uL 1-10 uL 10-100 uL 20-200 uL 100-1000uL

a. 525 uL b. 23.0 uL c. 1234 uL d. 3.55 uL

0525 0230 1234 0355

100-1000uL 10-100 uL 2x 100-1000uL 1-5uL

4) Sometimes scientists develop slang. We will call micropipettes P-highest number of range. Practice this lingo by
determining the amounts below.

19.5 125 25
5) Keep on practicing!

a. 95 b. 0.5
c. 35 d. 2.5