LABORATORY 1 TOOLS OF THE TRADE LSSI Alum
LABORATORY 1: TOOLS OF THE TRADE LSSI Alum Mary Haus, Chaffey Joint Union High School District
Curriculum & Standards • NGSS –HS-LS 1 -1 - Construct an explanation based on evidence for DNA, etc. –Science Practices - Asking Questions and Defining problems –DCI - LS 1. A - Structure and Function (both) –DCI - LS 1. C - Organization for Matter and Energy Flow in Organisms –DCI - LS 3. A - Inheritance of Traits www. amgenbiotechexperience. com
Curriculum & Standards • Common Core English Language Arts –RST. 11 -12. 9: Synthesize information from a range of sources into a coherent understanding… –WHST. 9 -12. 9: Draw evidence from informational texts to support analysis, reflection, and research. www. amgenbiotechexperience. com
Curriculum & Standards • 21 st Century Core Competencies - –Cognitive Domain - Cognitive Processes and Strategies: • Reasoning/argumentation • interpretation –Cognitive Domain - Knowledge: • Oral and written communication • Active listening –Interpersonal Domain - Teamwork and Collaboration: • Communication • Collaboration • Teamwork • Interpersonal Skills www. amgenbiotechexperience. com
Curriculum & Standards • Review pages in Teacher Guide: pages 49 -50 and 63 • Review Student Guide: Read page 19 www. amgenbiotechexperience. com
Lab Prep & Aliquouting Guidelines Reagents/Supplies 12 mls of Red Dye (RD)/kit 6 L of 1 x SB buffer/kit 6 mls Solution 1 (S 1) 6 mls Solution 2 (S 2) 6 mls Solution 3 (S 3) Micropipette Practice Sheet 11/kit Round Practice Plates 12/kit Each plate will have ~15 wells. Each student should practice pipetting into one well. 10 gels per class. If you prefer to have your students pour their own gels, please let us know when you make your reservation on LARS. Aliquot 1 ml/group . 5 ml/group N/A Storage Temp RT RT RT 4 o Notes Please return tubes Please return Bottles Please return tubes Please return sheets Please return used and unused plates N/A 4 o Please return any unused gels. Discard used gels in the trash. Equipment/Supplies 10 Student boxes with the following: 1 p 20 micropipette 1 microfuge rack 1 p 200 micropipette 1 bag of microfuge tubes 1 p 1000 micropipette 1 bag of microfuge tubes 1 waste and 1 box of refillable tips (2 ul-200 ul) 1 ice bucket 4 Mini centrifuges 10 Gel Electrophoresis chambers 10 Gel trays 10 gel combs 5 Power supplies www. amgenbiotechexperience. com
Safety General Lab Safety Guidelines • Use laboratory coats, safety glasses and gloves as appropriate • Avoid restrictive clothing and open-toed shoes • No eating or drinking in the lab • Make sure that students are familiar with the operating instructions and safety precautions before they use any of the lab equipment • Check all MSDS (Material Safety Data Sheets) for all chemicals and reagents in the lab before preparing and running the lab • Wash hands at the conclusion of the lab. Lab Safety Guidelines for lab 1 • Use caution with hot liquids and glassware. Wear heat-proof gloves to move hot glassware and liquids • Make sure the power is turned off on power supplies before connecting electrodes • After gel run is complete, turn off power supply then unplug electrodes • If the power is on, do not touch the buffer or electrophoresis equipment as you may receive an electrical shock • Never leave the electrophoresis power unit on without supervision. There is a risk of fire if the buffer leaks out or if the buffer should evaporate completely during electrophoresis www. amgenbiotechexperience. com
Lab Overview/Introduction • Teacher Manual – pgs 49 – 50 • Student Manual – pg 19 www. amgenbiotechexperience. com
Lab 1. 1 – How to use a micropipette • Teacher Manual – Preparation on pgs 51 – 53 • Student Manual – pgs 21 - 25 The Basic Laboratory Tool: Micropipette www. amgenbiotechexperience. com
An Introduction to Pipetting http: //www. teachersdomain. org/resource/biot 11. sci. life. gen. usingmicro/
Using a Micropipette Activity • Handout found on MATSC www. amgenbiotechexperience. com
Steps for using a Micropipette 1. Set the volume on the micropipette. 2. Use the micropipette to attach a pipette tip. 3. Pick up and hold the microcentrifuge tube containing the solution at eye-level 4. Push the plunger to the first stop. 5. Insert the pipette tip into the tube containing a solution. 6. Aspirate the solution by slowly releasing the plunger of the micropipette. 7. Dispense the solution by slowly pushing the plunger to the first stop and then to second stop. www. amgenbiotechexperience. com
Setting the Volume on a Micropipette Activity (found on MATSC) 1) The smallest amount of volume that I can measure with a P-20 pipette is… 2) When I use a P-20 pipette the maximum volume I can measure is… www. amgenbiotechexperience. com
Remember, The Pipette can be damaged if you pass these… •
Lab 1. 1: How to use a micropipette 23 Student Manual – pgs. 21 - 23 www. amgenbiotechexperience. com
Lab 1. 2 (part A) ● Practice loading into gel. ● Student manual pgs. 28 -29 ***Make sure to fill with 1 X SB buffer before loading. www. amgenbiotechexperience. com
Intro to Flowgramming • See additional power point www. amgenbiotechexperience. com
Learning a New Skill: Electrophoresis and Gel Analysis https: //www. youtube. com/watch? v=KDC 0 tl. AZH 9 k www. amgenbiotechexperience. com
Let’s Flowgram! ● ● ● In groups of 4 Use large whiteboards and markers Student manual, pgs. 29 - 31 Lab 1. 2 part B Lets compare : ) - teacher manual pg. 61 Now on to the lab! www. amgenbiotechexperience. com
Lab 1. 2 (Part B) ● Student Manual pgs. 26 - 32 ● Run three samples marked S 1, S 2, and S 3. (part B) ○ Solution Tube 1 = blue + purple ○ Solution. Tube 2 = blue + purple + yellow ○ Solution. Tube 3 = blue ● Yellow dye = orange G ● Purple dye = bromophenol blue ● Blue dye = xylene cyanole www. amgenbiotechexperience. com
Lab 1. 2 (Part B) 3 A 2 1 Which dye do you predict has the largest molecular weight? Remember – Things separate by size AND by charge (size is the biggest factor) www. amgenbiotechexperience. com
Some GE tips… • Gel box lids slide off, they do not pull off • Make sure that you pour enough buffer to completely cover the gel by about 2 mm. If you can see dimples in the buffer where the wells are you need to add more. • Wells should be on the black side so your samples can “RUN to RED. ” • Place your gel box close enough to connect to power supply so that you do not need to move it to plug it in. www. amgenbiotechexperience. com
While Samples are running… • Gel electrophoresis Card Sort • Gel electrophoresis Activity • Lab 1. 3 www. amgenbiotechexperience. com
Gel Electrophoresis Card Sort
Gel Electrophoresis Activity
Gel Electrophoresis Activity
Lab 1. 3 ● Optional Activity to demonstrate precision ● Found in teacher manual, pgs. 64 - 66 and RM 1 (last section of the manual) ● Complete if time permits
Lab 1. 2 – Results Orange G (yellow), mol. Wt. = 452. 4 Bromophenol blue (purple) = 691. 9 Xylene cyanole (blue) = 538. 6 We only use negatively charged dyes because DNA is negatively charged. Note that there are many bromine groups (which are K. Schramm negatively charged) on Bromphenol blue.
Lab 1. 2 Results S 1 S 2 S 3 Bromophenol blue (purple) is more negatively charged than xylene cyanole (blue) due to negatively charged bromine ions. Therefore, it travels farther than the smaller xylene cyanole. Don’t forget to get a photo of your gel! www. amgenbiotechexperience. com
Tips/Strategies/Questions • Preparation - • Pipette practice additional techniques – page covers, ziplocs • Results on overhead transparencies • Flowgrams on colored sheets by day • Who Dunnit? for different level classes www. amgenbiotechexperience. com
Contact Mary Haus Los Osos High School 6001 Milliken Ave. Rancho Cucamonga, CA 91737 909 -477 -6900 Mary. haus@cjuhsd. net www. amgenbiotechexperience. com
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