[Music]
[Music]

Welcome to Addgene's Intro to the Lab Bench!
Welcome to Addgene's Intro to the Lab Bench!

I'm Diego,
I'm Diego,

and I'm here to talk about one of the most important tools
and I'm here to talk about one of the most important tools

you'll encounter in your lab work...
you'll encounter in your lab work...

a pipette.
a pipette.

[Intro Music playing]
[Intro Music playing]

This small object is an essential tool
This small object is an essential tool

for nearly everyone in molecular biology.
for nearly everyone in molecular biology.

Using a pipette, you’ll be able to accurately dispense
Using a pipette, you’ll be able to accurately dispense

small amounts of liquid,
small amounts of liquid,

amounts between point one microliters, and 1 milliliter,
amounts between point one microliters, and 1 milliliter,

far too small to eyeball accurately.
far too small to eyeball accurately.

Your lab work needs to be precise,
Your lab work needs to be precise,

and any changes, even very small ones,
and any changes, even very small ones,

can negatively impact your experiment results.
can negatively impact your experiment results.

This video will teach you proper pipetting technique,
This video will teach you proper pipetting technique,

and how to select the proper pipette and tip for the job.
and how to select the proper pipette and tip for the job.

So, let’s get into it!
So, let’s get into it!

First, gather up your materials.
First, gather up your materials.

You'll need...
You'll need...

Pipette tips...
Pipette tips...

A waster container...
A waster container...

Containers to hold your measured liquid,
Containers to hold your measured liquid,

like a microcentrifuge tube, a bottle, or something else...
like a microcentrifuge tube, a bottle, or something else...

Liquid to pipette.
Liquid to pipette.

For this demonstration, we’ll just be using water.
For this demonstration, we’ll just be using water.

Something to label your containers...
Something to label your containers...

[marker squeaking]
[marker squeaking]

And of course.,
And of course.,

a pipette!
a pipette!

What kind of pipette you will need
What kind of pipette you will need

depends on your experiment.
depends on your experiment.

This video will only be covering single channel pipettes.
This video will only be covering single channel pipettes.

If you’re looking to improve
If you’re looking to improve

your multi-channel pipetting technique,
your multi-channel pipetting technique,

check out Addgene’s protocol video
check out Addgene’s protocol video

on multi-channel pipettes.
on multi-channel pipettes.

Back to single channel,
Back to single channel,

you’ll want to select the pipette based on the amount
you’ll want to select the pipette based on the amount

of liquid you’ll be pipetting.
of liquid you’ll be pipetting.

There are five standard sizes to choose from:
There are five standard sizes to choose from:

P2...
P2...

P10...
P10...

P20...
P20...

P100...
P100...

P200...
P200...

and P1000.
and P1000.

Have your pipette?
Have your pipette?

Great!
Great!

That means we it's time to talk about
That means we it's time to talk about

the anatomy of a pipette.
the anatomy of a pipette.

[Music]
[Music]

Here's a P1000 pipette.
Here's a P1000 pipette.

From the front, you can see the plunger at the top.
From the front, you can see the plunger at the top.

The plunger is used to both draw liquid into
The plunger is used to both draw liquid into

and expel it from the pipette tip.
and expel it from the pipette tip.

Next, in the middle, you can see the volume display.
Next, in the middle, you can see the volume display.

This shows the set volume amount
This shows the set volume amount

that will be drawn up into the tip.
that will be drawn up into the tip.

We’ll show you how to read this display a little later.
We’ll show you how to read this display a little later.

Finally, we have the tip ejector ring.
Finally, we have the tip ejector ring.

This pushes the pipette tip off the pipette,
This pushes the pipette tip off the pipette,

allowing you to quickly dispose of used tips.
allowing you to quickly dispose of used tips.

We’ll also explain why that’s important in a bit.
We’ll also explain why that’s important in a bit.

Turning the pipette on its side,
Turning the pipette on its side,

you’ll see the volume adjustment ring.
you’ll see the volume adjustment ring.

This is what you’ll use to set the volume
This is what you’ll use to set the volume

of liquid the tip will hold.
of liquid the tip will hold.

Next you’ll see the tip ejector button,
Next you’ll see the tip ejector button,

and further down the tip ejector itself.
and further down the tip ejector itself.

This is the mechanism that physically pushes the tip
This is the mechanism that physically pushes the tip

away from the pipette, for those easy tip disposals.
away from the pipette, for those easy tip disposals.

Feeling more comfortable with the pipette?
Feeling more comfortable with the pipette?

Great!
Great!

Let’s talk about setting and reading the volume
Let’s talk about setting and reading the volume

on the pipette.
on the pipette.

Setting the volume is the same across all standard pipettes.
Setting the volume is the same across all standard pipettes.

Just rotate the ring below the plunger to increase
Just rotate the ring below the plunger to increase

or decrease the volume.
or decrease the volume.

But,
But,

reading the display is slightly different
reading the display is slightly different

across different pipette sizes.
across different pipette sizes.

For a P1000 pipette,
For a P1000 pipette,

the first red digit is thousands of microliters,
the first red digit is thousands of microliters,

the middle is hundreds,
the middle is hundreds,

and the third is tens.
and the third is tens.

So, this is 1,000 microliters.
So, this is 1,000 microliters.

While this would be 650 microliters.
While this would be 650 microliters.

For the P200,
For the P200,

he first digit is hundreds,
he first digit is hundreds,

the second is tens,
the second is tens,

and the third ones.
and the third ones.

So, one-zero-zero
So, one-zero-zero

from top to bottom, would be 100 microliters,
from top to bottom, would be 100 microliters,

while zero-nine-five
while zero-nine-five

from top to bottom would be 95 microliters.
from top to bottom would be 95 microliters.

The P20,
The P20,

P10,
P10,

and P2 all follow similar patterns.
and P2 all follow similar patterns.

Keep in mind that this dictates what tips you’ll be using.
Keep in mind that this dictates what tips you’ll be using.

Different tips hold different volumes of liquid.
Different tips hold different volumes of liquid.

Check the information on the box
Check the information on the box

find out the volume range for each tip.
find out the volume range for each tip.

Now that you’re familiar with the pipette and tips,
Now that you’re familiar with the pipette and tips,

let’s talk about using the pipette.
let’s talk about using the pipette.

First, set the desired volume of the liquid
First, set the desired volume of the liquid

using the adjustment ring.
using the adjustment ring.

It’s often standard practice to go above,
It’s often standard practice to go above,

then roll the dial back down the desired volume.
then roll the dial back down the desired volume.

On some pipettes,
On some pipettes,

this can be achieved by rotating the dials
this can be achieved by rotating the dials

behind the numerical display.
behind the numerical display.

To start pipetting, you’ll want to hold the pipette like this,
To start pipetting, you’ll want to hold the pipette like this,

with your index finger below the notch,
with your index finger below the notch,

and your thumb on the plunger like so.
and your thumb on the plunger like so.

Next, select the pipette tip designed to fit your pipette.
Next, select the pipette tip designed to fit your pipette.

To load the tip,
To load the tip,

open the tip box,
open the tip box,

and push the end of the pipette onto the tip.
and push the end of the pipette onto the tip.

Before you move on,
Before you move on,

ensure the tip is properly set in the pipette.
ensure the tip is properly set in the pipette.

Do not touch the tips with your fingers.
Do not touch the tips with your fingers.

Doing so could contaminate the tips,
Doing so could contaminate the tips,

or puncture the tip of your glove.
or puncture the tip of your glove.

Now, gently push down the plunger.
Now, gently push down the plunger.

You may notice several “stops,”
You may notice several “stops,”

where you feel resistance while pressing down.
where you feel resistance while pressing down.

Stop at that first “stop,”
Stop at that first “stop,”

when you first start to feel that resistance.
when you first start to feel that resistance.

This expels the exact amount of air from the tip
This expels the exact amount of air from the tip

equal to the amount of liquid you previously set.
equal to the amount of liquid you previously set.

Keep the plunger held down,
Keep the plunger held down,

and gently lower the tip into the liquid.
and gently lower the tip into the liquid.

[liquid pouring]
[liquid pouring]

Do not submerge the entire pipette.
Do not submerge the entire pipette.

Only the tip should ever be submerged into the liquid.
Only the tip should ever be submerged into the liquid.

Once the tip is submerged,
Once the tip is submerged,

slowly release the plunger,
slowly release the plunger,

allowing it to go back to its resting position.
allowing it to go back to its resting position.

Wait until the liquid has finished flowing into the pipette tip.
Wait until the liquid has finished flowing into the pipette tip.

Keep in mind, the more viscous the liquid,
Keep in mind, the more viscous the liquid,

the lower the flow rate into the pipette tip.
the lower the flow rate into the pipette tip.

Here's a pro tip!
Here's a pro tip!

If you’re pipetting from a relatively large container
If you’re pipetting from a relatively large container

holding a small amount of liquid,
holding a small amount of liquid,

tilt the container so it’s easier to reach the liquid.
tilt the container so it’s easier to reach the liquid.

Once the liquid has finished drawing into the pipette tip,
Once the liquid has finished drawing into the pipette tip,

lift the pipette so that the tip exits the liquid.
lift the pipette so that the tip exits the liquid.

When you’re lifting the pipette tip from the liquid,
When you’re lifting the pipette tip from the liquid,

no liquid should drip from the tip.
no liquid should drip from the tip.

If it does, this could indicate
If it does, this could indicate

the tip is not properly seated on the pipette.
the tip is not properly seated on the pipette.

Now,
Now,

place the pipette tip into the container you're using
place the pipette tip into the container you're using

to hold the measured liquid.
to hold the measured liquid.

This is your microcentrifuge tube, conical tube, or plate.
This is your microcentrifuge tube, conical tube, or plate.

If the tube already has liquid,
If the tube already has liquid,

insert the tip into the liquid.
insert the tip into the liquid.

Now, slowly press down the plunger to the first stop.
Now, slowly press down the plunger to the first stop.

Wait,
Wait,

then press down all the way to the second stop.
then press down all the way to the second stop.

This ensures that you’ll expel any excess liquid from the tip.
This ensures that you’ll expel any excess liquid from the tip.

If this is a new container,
If this is a new container,

Place the tip lightly against the side of the tube.
Place the tip lightly against the side of the tube.

Capillary action between the side of the tube and the liquid
Capillary action between the side of the tube and the liquid

Will help draw all of the liquid out of the tip.
Will help draw all of the liquid out of the tip.

Now, lift the pipette tip completely out of the liquid,
Now, lift the pipette tip completely out of the liquid,

and relase the plunger,
and relase the plunger,

being careful to not touch the sides of the container
being careful to not touch the sides of the container

with the pipette.
with the pipette.

Discard the pipette tip by holding the tip over
Discard the pipette tip by holding the tip over

your dedicated waste container
your dedicated waste container

and pressing the tip ejector button.
and pressing the tip ejector button.

If you’re pipetting biohazardous material,
If you’re pipetting biohazardous material,

make sure to discard the tip into a biohazardous waste container.
make sure to discard the tip into a biohazardous waste container.

Remember, never reuse pipette tips.
Remember, never reuse pipette tips.

Reusing tips could lead to contamination
Reusing tips could lead to contamination

or inaccurate liquid amounts.
or inaccurate liquid amounts.

Always use a fresh tip when you’re pipetting liquid.
Always use a fresh tip when you’re pipetting liquid.

If you're struggling with hand steadiness
If you're struggling with hand steadiness

or accuracy,
or accuracy,

try putting two fingers on the pipette for support.
try putting two fingers on the pipette for support.

You can also rest the elbow of your pipetting arm
You can also rest the elbow of your pipetting arm

on the bench for added stability.
on the bench for added stability.

Congratulations! You just successfully used a pipette!
Congratulations! You just successfully used a pipette!

If you had any trouble, don’t worry.
If you had any trouble, don’t worry.

Trust me, you’ll get plenty of practice in the lab.
Trust me, you’ll get plenty of practice in the lab.

As mentioned before, if you’re pipetting the same amount
As mentioned before, if you’re pipetting the same amount

of liquid into different tubes or into the wells of a microplate,
of liquid into different tubes or into the wells of a microplate,

you may want to use a multichannel pipette.
you may want to use a multichannel pipette.

Check out Addgene’s video on mulitchannel pipettes
Check out Addgene’s video on mulitchannel pipettes

to learn more.
to learn more.

Thanks for watching another Addgene Intro to the Lab Bench video.
Thanks for watching another Addgene Intro to the Lab Bench video.

Make sure you leave a comment down below,
Make sure you leave a comment down below,

tell us how you like the video,
tell us how you like the video,

or let us know what topic you’d most like to see us cover.
or let us know what topic you’d most like to see us cover.

Addgene: a better way to share science.
Addgene: a better way to share science.