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Bellwork

Bellwork. What is an SOP? What is its purpose? Why is Quality Control important?. How to Properly Use a Micropipettor. Bioscience : Quality Control. Objectives. Identify the different sizes of micropipettors. Recognize when to use which size micropipettor.

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Bellwork

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  1. Bellwork • What is an SOP? • What is its purpose? • Why is Quality Control important? Property of CTE Joint Venture

  2. How to Properly Use a Micropipettor Bioscience: Quality Control

  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. Property of CTE Joint Venture

  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. Property of CTE Joint Venture

  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. Property of CTE Joint Venture

  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 Property of CTE Joint Venture

  7. Write onyour Slates: Which micropipettor will you use for the following volumes? 254 uL 7 uL 98 uL 645 uL 145 uL 2 PROPERTY OF PIMA COUNTY JTED, 2010

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

  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 Property of CTE Joint Venture

  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? Property of CTE Joint Venture

  11. Micropipettor Use – Demo Video http://www.youtube.com/watch?v=wlO4zJLR8R8&feature=related8/14/2014 How to pipet http://www.youtube.com/watch?v=nv5Bi2Wy1KU&feature=related Common errors Property of CTE Joint Venture

  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 Property of CTE Joint Venture

  13. Micropipettor Use - DON’Ts • http://www.youtube.com/watch?v=1J3w4cS2MvM • 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 Property of CTE Joint Venture

  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. Property of CTE Joint Venture

  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.

  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

  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. Property of CTE Joint Venture

  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. Property of CTE Joint Venture

  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. Property of CTE Joint Venture

  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. Property of CTE Joint Venture

  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. Property of CTE Joint Venture

  22. 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 . . . 22 PROPERTY OF PIMA COUNTY JTED, 2010

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