1. Bellwork What is an SOP? What is its purpose?
Why is Quality Control important?
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2. How to Properly Use a Micropipettor
Bioscience: Quality Control
How to Properly Use a Micropipettor
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3. Objectives Identify the different sizes of micropipettors.
Recognize when to use which size micropipettor.
Demonstrate ability to accurately set and use the different sized micropipettors.
Demonstrate ability to successfully complete Micropipetting SOP.
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4. Purpose Introduce the laboratory instrument: micropipettor.
Practice using different sized micropipettors to gain accuracy and precision.
Introduce the process of correctly following an SOP.
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5. What are Micropipettors?!
We will be using 3 different sizes of micropipettors:
P20
P200
P1000
Very expensive and very fragile – USE Responsibly!!!
Used to measure liquid amounts LESS THAN or EQUAL to 1 mL.
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6. When to Use Each Micropipettor
Each micropipettor has its name on the top cap.
Each micropipette has a limit:
P20: 0.5 to 20.0 uL
P200: 20 to 200 uL
P1000: 200 to 1000 uL
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7. PROPERTY OF PIMA COUNTY JTED, 2010
Write on your Slates:
Which micropipettor will you use for the following volumes? 254 uL 7 uL 98 uL 645 uL 145 uL
2. Whiteboards/Slates
student writes answer or solution to a question posed by the teacher
teacher solicits all students to show at the same time
teacher gives feedback to the students
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8. How to Set/Adjust the Micropipettor
Adjustment knobs are on the side of each micropipettor.
Red digits indicate a decimal place
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9. How to Load and Dispense Liquids
Place clean plastic tip at end of micropipettor
Plunger has TWO different ‘stop’ positions.
First is used to load (pick up) liquid
The second (lower) is used to dispense (expel) liquid
Two stop method is what creates accuracy and precision of instrument – YOU MUST BE CONSISTENT
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10. Micropipettor Use – Video Questions
While watching video answer the following 3 questions:
How does the scientist know that she has the correct micropipettor?
Why do you use the first stop on the plunger?
How do you dispense liquid?
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11. Micropipettor Use – Demo Video
How to pipet
Common errors
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12. Micropipettor Use - DOs
Always use the correct micropipettor for the volume being measured
Always use the correct pipette tip for each size micropipettor
Use the first stop for loading
Use the second stop for dispensing
Release plunger slowly
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13. Micropipettor Use - DON’Ts
S2MvM
NO Double Dipping
When in doubt – CHANGE TIPS!!
Don’t place tip into liquid too deeply.
Use the two stops appropriately
Don’t squirt liquid out too quickly
Don’t point tip up – this will contaminate the barrel
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14. Partner Practice
Work with a partner to create a list of 5 things you should check before using your micropipettors.
Use your slates to make a list. Be ready to be called on.
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15. Micropipettor Lab You will be working in pairs:
Who will be leader/implementer?
set-up/clean-up?
Both people in your group need to complete the pipetting portion of this lab.
Write down materials and procedures into your laboratory notes.
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16. Micropipettor Lab: Materials
For each pair you need:
Two micropipettors (P20, P200)
One box of yellow tips
Two microtiter plates
One set of dyes (red, yellow, blue)
One 50 mL beaker of water
Two plastic sheet protectors
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17. Micropipettor Lab: Results
You will use a variable micropipette to prepare seven different dye mixtures in the wells of a microtiter plate.
You will then dispense 5 μl of each of the seven diluted dye solutions onto a pipet card in triplicate.
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18. Micropipettor Procedures
Gather materials
Label a microtiter plate with your initials and label seven of the wells as number 1 – 7.
MAKE SURE TO CHANGE TIPS INBETWEEN EACH DISPENSE.
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19. Micropipettor Lab: Procedures (cont.)
Prepare the dye solutions, one per well, using the ingredients shown in the following table.
You will fill in LAST THREE columns.
Well
Red (uL)
Blue (uL)
Yellow (uL)
Water (uL)
Total (uL)
Resulting Color
Under, over, correct? 1 5 - 40 2 10 35 3 25 4 15 13 32 6 30 7 33
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20. Micropipettor Lab: Verification
Once solutions are made, set your micropipettor to the Total Volume you determined and check the volume in the well in each well.
If you under pipetted, you will see air at the end of the tip.
If you over pipetted you will have liquid left in the tube.
Record results on chart.
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21. Micropipettor Lab: Consistency/Precision
Once you have checked your pipetting accuracy using the micropipette, practice dispensing liquids by placing 5 uL of each dye onto a plastic sheet protector in triplicate.
You will have 7 rows of three drops neatly arranged on your sheet protector when complete.
Have your teacher sign-off your lab results and then complete closure.
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22. PROPERTY OF PIMA COUNTY JTED, 2010
At the end of lab write up –Complete the 4 Statements
I would like to know
more about . . .
I now
understand . . .
I need more
help
with . . .
I could
teach
someone
else to . . .
 23. Outcome Statements
teacher gives the following prompts
I now understand . . .
I would like to know more about . . .
I need more help with . . .
I could teach someone else to . . .
teacher collects their written responses or does a think-pair-share
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