Bellwork What is an SOP? What is its purpose? Why is Quality Control important? 4/15/2017 Property of CTE Joint Venture

How to Properly Use a Micropipettor Bioscience: Quality Control How to Properly Use a Micropipettor 4/15/2017

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. 4/15/2017 Property of CTE Joint Venture

Purpose Introduce the laboratory instrument: micropipettor. Practice using different sized micropipettors to gain accuracy and precision. Introduce the process of correctly following an SOP. 4/15/2017 Property of CTE Joint Venture

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. 4/15/2017 Property of CTE Joint Venture

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 4/15/2017 Property of CTE Joint Venture

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 PROPERTY OF PIMA COUNTY JTED, 2010 2

How to Set/Adjust the Micropipettor Adjustment knobs are on the side of each micropipettor. Red digits indicate a decimal place 4/15/2017 Property of CTE Joint Venture

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 4/15/2017 Property of CTE Joint Venture

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? 4/15/2017 Property of CTE Joint Venture

Micropipettor Use – Demo Video http://www.youtube.com/watch?v=wlO4zJLR8R8&feature=related4/15/20174/15/2017 How to pipet http://www.youtube.com/watch?v=nv5Bi2Wy1KU&feature=related Common errors Property of CTE Joint Venture

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 4/15/2017 Property of CTE Joint Venture

Micropipettor Use - DON’Ts http://www.youtube.com/watch?v=1J3w4c 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 4/15/2017 Property of CTE Joint Venture

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. 4/15/2017 Property of CTE Joint Venture

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. 4/15/2017

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 4/15/2017

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. 4/15/2017 Property of CTE Joint Venture

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. 4/15/2017 Property of CTE Joint Venture

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 4/15/2017 Property of CTE Joint Venture

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. 4/15/2017 Property of CTE Joint Venture

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. 4/15/2017 Property of CTE Joint Venture

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 PROPERTY OF PIMA COUNTY JTED, 2010 22