Bio-Rad Biotechnology Explorer IDEA and STEM Kits Pop Culture Science Bio-Rad Biotechnology Explorer IDEA and STEM Kits

Curriculum and Training Specialists Damon Tighe damon_tighe@bio-rad.com Sherri Andrews, Ph.D. sherri_andrews@bio-rad.com Leigh Brown leigh_brown@bio-rad.com

Bio-Rad Resources @BioRadEducation Biotechnology: A Lab Skills Course Pinterest.com/teachbiology bit.ly/b-rdropbox

Workshop Overview What are we doing? Pop Culture + Lab Techniques How do we get students engaged? Movie scenarios Deeper dive into technique Choose your favorite movie scenario! Lab tailored to movie pick Standards alignment Other ideas? Results

Getting Students Engaged

A Few of your Students’ Favorite Movies? Interstellar DESPICABLE ME 2 DIVERGENT

Electrophoresis separates molecules by CHARGE and SIZE Molecular sieve Electricity Electrophoresis: “carry with water”

Lapse It! (10 second intervals)

Interstellar Interstellar Link to trailer of Interstellar movie: https://www.youtube.com/watch?v=2LqzF5WauAw

Insterstellar https://www.youtube.com/watch?v=zSWdZVtXT7E

Interstellar Setting the scene: Crop blight has caused a massive die-off of most food crops. You are in a crew of astronauts who travel through a wormhole, searching for a new planetary home for humanity with a suitable food source.

Interstellar Premise: You find some potentially edible specimens from an exoplanet near the wormhole, but some are poisonous, and contain a visible blue dye. Students must test the potential food source to find out if it contains the poisonous dye.

potential food sources Interstellar Lab Set Up: Each student has several specimens (M&Ms candy) to test. They use gel electrophoresis to determine if the blue dye is any of the specimens. poisonous blue dye potential food sources Teacher Prep: Blue 1 FD&C dye used for a standard (to represent poisonious blue dye) Medicine cup full of M&Ms or other candies (Nerds, Skittles) – students can separate colors out into other cups.

Link to trailer of Divergent movie: http://divergentthemovie.com/

Divergent https://www.youtube.com/watch?v=336qJITnDi0

Divergent Setting the scene: You are a teenager in post-apocalyptic Chicago. Everyone is divided into 5 factions based on their aptitude and personality.

Divergent Premise: During your 16th year, you must decide whether to stay in your family’s faction, or join a different faction and leave your family behind. There are several ways to determine which faction you belong in, including an aptitude test.

Divergent After your aptitude test, results are now confirmed using a new technique to see what faction you most closely resemble. Do you fit in one faction? Or are you divergent, showing aptitude for 2 or more factions? **Results for Candor may be inconclusive. Hint: You might want to set out bowls of different colors of candies here for a visual for your students

Divergent Teacher Prep: Lab Set Up: Students will prepare extracts from the “faction samples” (candy). You might want to have these separated out into colors at the front of the room, each in its own bowl. These extracts will be run on a gel, along with 3 student samples. Students will identify their sample as matching one faction, or multiple ones (i.e. divergent), and decide if additional tests are needed or if the results are conclusive. Teacher Prep: M&Ms or Skittles (5 different colors) to represent each faction FD&C dyes for student samples – some should be a mix of 2 or more colors. Others should just be 1 color. Mix 1:1. faction samples student sample A student sample B student sample C

DESPICABLE ME 2 Despicable Me 2 Link to trailer of Despicable Me 2 movie: https://www.youtube.com/watch?v=u6RxZK3LTnA

Despicable Me 2 Setting the scene: PX-41 is a dangerous mutagen engineered by villians. It can turn minons into furry killing machines, capable of destroying everything in their paths. You are a reformed evil scientist, looking for an antidote to cure the mutated minions.

Despicable Me 2 Premise: Using the materials available to you in your not-so-evil scientist laboratory, hatch a plan to purify different antidotes. You will need to: Identify which samples contain the yellow antidote. Are any of them pure? Can you find another source for the antidote?

Potential antidote sources (students pick 2 colors to test) Despicable Me 2 Lab Set Up: Each workstation has a different set of potential antidotes, made from FD&C dyes. One is pure (yellow) – the others are mixes of various dyes. Teacher Prep: FD&C dye – yellow, plus mixes of 2 or more other FD&C dyes (include yellow in a few) Optional – M&Ms or Skittles for students to test for the presence of an antidote. pure antidote sample Sample A Sample B Sample C Sample D Sample E Potential antidote sources (students pick 2 colors to test)

Before we start…a little background What is electrophoresis?

How can you separate molecules?

Insert close up of FD&C dyes in kit Food and marker dyes have an intrinsic SIZE and CHARGE and thus can be separated using Electrophoresis Food dye Molecular weight Charge FD&C Blue 1 792.86 -2 FD&C Yellow 5 534.37 -3 FD&C Yellow 6 452.37 FD&C Red 40 496.43 Carminic Acid 492.38 Beetroot red 551.48 +1 Insert close up of FD&C dyes in kit

DIY vs Commercial Electrophoresis Chamber

Interstellar Procedure - Interstellar Time to Pick! Interstellar Procedure - Interstellar Testing potential food source samples for poisonous blue dye Divergent Testing student samples to determine which faction(s) they belong to Despicable Me Testing samples to determine which ones contain the antidote, and if any are pure

Procedure - Interstellar Before we begin: Find the Blue 1 tube in your green rack. This will represent the poisonous blue dye that we are testing for. Locate the potential food sources (M&Ms). Find the Extraction Buffer tubes. We won’t be using any of the other tubes in the rack, so you can put them to the side. poisonous blue dye potential food sources Extraction buffer

Procedure - Interstellar (Already done for you). Place comb across box at 3cm mark. Pour molten agarose into box. Let solidify. Remove comb. Use a plastic ruler or card to cut across gel about 1 cm from each end, and scoop out and discard the rectangle of agarose. Center gel in plastic chamber so that the wells are going across the box.

Procedure - Interstellar Add enough TAE buffer to completely cover gel. Unfold paper clip electrodes as shown in diagram. Place one at each end of the gel. The bottom of the “U” will rest in the empty space at the top and bottom of the gel.

Procedure - Interstellar Separate M&Ms by color. Put one candy of each color into each cup. Add 250 ul of extraction buffer to each cup. Swirl cups so dyes come off into the buffer. Extraction buffer 250 ul

Procedure - Interstellar Use the green 10 ul fixed volume pipet to load 10 ul of each of your samples into the wells. Change tips between each sample. Lane 1: Blue 1 (poisonous dye) Lane 2: Yellow M&M Lane 3: Red M&M Lane 4: Brown M&M Lane 5: Blue M&M Lane 6: Orange M&M Lane 7: Green M&M ***note: order is not important – just be sure to write down what you did! Load 10 ul of each samplet

Procedure - Interstellar Assemble your battery tower as shown in the diagram. Use the black alligator clip to clip your paper clip to the container at the end closest to your sample wells. Use the red alligator clip to attach the paper clip at the other end of the gel. Catching both the container and the paper clip with the alligator clip ensures that the paper clip won’t fall over.

Procedure - Interstellar Start your gel by clipping the free end of the black alligator lead to the (-) terminal of your battery tower. Attach the free end of the red alligator lead to the (+) terminal of the battery tower. Look for bubbles. Let your gel run for about 15-20 minutes. Compare the Blue #1 standard to your food samples, and see if any contain the poisonous blue dye.

Before we begin Procedure - Divergent Remove any green M&Ms from your candy cup – we won’t need them during the lab. The other colors will represent each faction. Pick one sample (A, B, C, D, or E) for each person at your workstation. Find the Extraction Buffer tubes. We won’t be using any of the other tubes in the rack, so you can put them to the side. faction samples student sample A student sample B student sample C Extraction buffer

Procedure - Divergent (Already done for you). Place comb across box at 3cm mark. Pour molten agarose into box. Let solidify. Remove comb. Use a plastic ruler or card to cut across gel about 1 cm from each end, and scoop out and discard the rectangle of agarose. Center gel in plastic chamber so that the wells are going across the box.

Procedure - Divergent Add enough TAE buffer to completely cover gel. Unfold paper clip electrodes as shown in diagram. Place one at each end of the gel. The bottom of the “U” will rest in the empty space at the top and bottom of the gel.

Procedure - Divergent You have representative samples from each faction in front of you. Separate these by color, putting one of each color into each cup. Add 250 ul of extraction buffer to each cup. Swirl cups so dyes come off into the buffer. Extraction buffer 250 ul

Procedure - Divergent Use the green 10 ul fixed volume pipet to load 10 ul of each of your samples into the wells. Change tips between each sample. Lane 1: Yellow (Abnegation) Lane 2: Red (Amity) Lane 3: Brown (Candor) Lane 4: Blue (Erudite) Lane 5: Orange (Dauntless) Lane 6: Student A sample Lane 7: Student B sample Lane 8: Student C sample Load 10 ul of each samplet

Procedure - Divergent Assemble your battery tower as shown in the diagram. Use the black alligator clip to clip your paper clip to the container at the end closest to your sample wells. Use the red alligator clip to attach the paper clip at the other end of the gel. Catching both the container and the paper clip with the alligator clip ensures that the paper clip won’t fall over.

Procedure - Divergent Start your gel by clipping the free end of the black alligator lead to the (-) terminal of your battery tower. Attach the free end of the red alligator lead to the (+) terminal of the battery tower. Look for bubbles. Let your gel run for about 15-20 minutes. Compare the student samples to the faction samples. Which faction do you belong to? Are you divergent?

Procedure – Despicable Me Before we begin Find the following tubes in your green rack: Yellow 5 (pure antidote sample) Samples A, B, C, D, and E from potential antidotes. Extraction Buffer We won’t be using any of the other tubes in the rack, so you can put them to the side. Decide which 2 colors of candies might be a potential source of the yellow antidote. Take one of each of those 2 colors out, and put the rest of the candies aside. pure antidote sample Sample A Sample B Sample C Sample D Sample E Extraction buffer Potential antidote sources (pick 2 colors to test)

Procedure – Despicable Me (Already done for you). Place comb across box at 3cm mark. Pour molten agarose into box. Let solidify. Remove comb. Use a plastic ruler or card to cut across gel about 1 cm from each end, and scoop out and discard the rectangle of agarose. Center gel in plastic chamber so that the wells are going across the box.

Procedure – Despicable Me Add enough TAE buffer to completely cover gel. Unfold paper clip electrodes as shown in diagram. Place one at each end of the gel. The bottom of the “U” will rest in the empty space at the top and bottom of the gel.

Procedure – Despicable Me You have samples which potentially contain the antidote sitting in front of you. You also have some purified antidote. If you’d like, you may test 2 M&M samples to see if they contain the antidote as well. Add 250 ul of extraction buffer to each cup. Swirl cups so dyes come off into the buffer. Extraction buffer 250 ul

Procedure – Despicable Me Use the green 10 ul fixed volume pipet to load 10 ul of each of your samples into the wells. Change tips between each sample. Lane 1: Yellow 5 (Antidote) Lane 2: A Lane 3: B Lane 4: C Lane 5: D Lane 6: E Lane 7: Yellow M&M Lane 8: Green M&M Load 10 ul of each samplet

Procedure – Despicable Me Assemble your battery tower as shown in the diagram. Use the black alligator clip to clip your paper clip to the container at the end closest to your sample wells. Use the red alligator clip to attach the paper clip at the other end of the gel. Catching both the container and the paper clip with the alligator clip ensures that the paper clip won’t fall over.

Procedure – Despicable Me Start your gel by clipping the free end of the black alligator lead to the (-) terminal of your battery tower. Attach the free end of the red alligator lead to the (+) terminal of the battery tower. Look for bubbles. Let your gel run for about 15-20 minutes. Compare the purified antidote sample in lane 1 to the potential antidote samples. Which ones contain the antidote? Are any pure?

What Can You Teach?

Guardians of the Galaxy Other Ideas? THE HUNGER GAMES Guardians of the Galaxy Jurassic World

Results - Interstellar Yellow M&M Red M&M Brown M&M Blue M&M Orange M&M Green M&M Blue # 1

Make new gel with Student A (yellow 6), B, E Results - Divergent Yellow (Abnegation) Orange (Dauntless) Brown (Candor) Red (Amity) Blue (Erudite) Student E Student B Student C Make new gel with Student A (yellow 6), B, E

Results – Despicable Me Antidote (Yellow 5) Sample D Sample B Sample A Sample C Sample E Yellow M&M Green M&M

Teacher Prep and Lab Setup – for all 3 scenarios Create the following samples to be used in Despicable Me or Divergent. Mixtures should be equal parts. A: Yellow 6 B: Yellow 5 + Yellow 6 C: Yellow 5 + Blue 1 + Red 40 D: Blue 1 + Yellow 6 E: Yellow 5 + Blue 1 You will also need the following FD&C dyes: For Interstellar: Blue 1 For Despicable Me: Yellow 5 Each station also should have: 6 medicine cups 1 cup with M&Ms (mixed colors) Micropipet and tips Dye extraction buffer (2 ml) Gel STEM gel box (box, paper clip, alligator clips, batteries) Sharpie

IDEA STEM Electrophoresis Kit IDEA/STEM Electrophoresis Kits Found in the 2014/2015 Catalog on pg. 48-49