WEBVTT 00:00:00.280 --> 00:00:14.863 [upbeat electronic music...] 00:00:45.132 --> 00:00:45.832 [music fades] 00:00:45.837 --> 00:00:50.150 PRESENTER: The materials you will need are one conventional, non-GM soybean. 00:00:50.150 --> 00:00:51.818 One GM soybean. 00:00:51.818 --> 00:00:56.022 Two reaction vials or one microliter micro tube. 00:00:56.256 --> 00:00:57.991 Two transfer pipettes. 00:00:57.991 --> 00:01:00.326 Two lateral flow test strips. 00:01:00.326 --> 00:01:03.329 1% PBS buffer or distilled water. 00:01:03.563 --> 00:01:05.131 Four weigh boats. 00:01:05.131 --> 00:01:10.103 Access to tweezers, scoopulas, paper towels, hammer, 00:01:10.103 --> 00:01:12.614 and isopropyl alcohol wipes. 00:01:12.872 --> 00:01:15.508 Using the seeds in the GM soybean seed kit, 00:01:15.508 --> 00:01:18.244 allow students to observe and compare the seeds. 00:01:18.244 --> 00:01:22.682 Students should look for observable differences in shape, size, and color. 00:01:23.016 --> 00:01:26.820 Note: Little to no difference will be observable in the seeds. 00:01:27.020 --> 00:01:28.421 As students observe, 00:01:28.421 --> 00:01:31.191 be sure that the two varieties are kept separate. 00:01:31.191 --> 00:01:34.260 Explain to the class that although the two seeds look the same, 00:01:34.527 --> 00:01:36.019 there is a difference between them 00:01:36.019 --> 00:01:38.898 and we are going to do an experiment to try and discover it. 00:01:38.932 --> 00:01:41.701 Complete the following lab tests for the class to see. 00:01:41.701 --> 00:01:44.871 Label the micro tubes for identification of the seed 00:01:44.871 --> 00:01:49.776 that will be tested by labeling one "A", and the other "B". 00:01:50.877 --> 00:01:53.780 Break the seed by placing one GM soybean 00:01:53.780 --> 00:01:57.150 between two small weight boats and tapping it with a hammer. 00:01:57.817 --> 00:02:00.320 The seed should break into 2 or 3 pieces 00:02:00.320 --> 00:02:03.456 to allow enough surface area to be exposed for extraction. 00:02:03.823 --> 00:02:06.092 Note: Do not crush the seed. 00:02:06.092 --> 00:02:09.690 Crushing can cause issues recovering all pieces for extraction 00:02:09.762 --> 00:02:12.932 and may cause cross-contamination of the testing area. 00:02:13.433 --> 00:02:16.035 Repeat this step with the conventional soybean 00:02:16.035 --> 00:02:18.168 using separate weigh boats. 00:02:19.172 --> 00:02:20.974 Remove the top weigh boats 00:02:20.974 --> 00:02:23.977 and place the seed pieces in the correct micro tube. 00:02:24.077 --> 00:02:27.572 If the seed is stuck to the boat, use tweezers or a scoopula 00:02:27.572 --> 00:02:28.882 to gently release it. 00:02:28.882 --> 00:02:32.685 Do not touch the seeds with your hands to avoid cross-contamination. 00:02:33.186 --> 00:02:37.973 Clean tweezers with 70% ethanol or isopropyl alcohol wipes 00:02:38.303 --> 00:02:40.160 to prevent cross-contamination. 00:02:40.693 --> 00:02:43.447 If any of the particles of the GM seed sample 00:02:43.447 --> 00:02:45.365 end up in the conventional sample, 00:02:45.365 --> 00:02:48.034 you will get a false positive on the test strip. 00:02:48.034 --> 00:02:50.281 So, it is important to use different weigh boats 00:02:50.281 --> 00:02:51.638 and to clean the scoopula 00:02:51.638 --> 00:02:54.641 to prevent what we call in the lab, "cross-contamination". 00:02:55.441 --> 00:03:01.011 Use the pipette to fill the micro tube with 1% PBS buffer or distilled water. 00:03:01.314 --> 00:03:03.746 Approximately 0.5 milliliters 00:03:04.083 --> 00:03:08.388 Using the pipette as a pestle and the micro tube as a mortar, 00:03:08.721 --> 00:03:12.592 stir in the seed pieces for about 20 to 30 seconds. 00:03:13.293 --> 00:03:16.829 Be sure to stir with separate pipettes to avoid cross-contamination. 00:03:17.263 --> 00:03:22.569 Let the micro tube with the pieces of seed and distilled water or 1% PBS buffer, 00:03:22.869 --> 00:03:25.505 stand for 3 to 5 minutes. 00:03:25.505 --> 00:03:28.652 Place one lateral flow test strip inside each vial 00:03:28.652 --> 00:03:30.785 with the arrow pointing downward. 00:03:31.377 --> 00:03:33.173 Allow the test to continue 00:03:33.173 --> 00:03:36.115 in the micro tube at room temperature for five minutes. 00:03:36.382 --> 00:03:39.704 During this time, explain how the lateral flow strip test works 00:03:39.704 --> 00:03:40.954 using the lesson plan. 00:03:41.721 --> 00:03:43.323 Interpreting the results. 00:03:43.323 --> 00:03:47.059 If the sample contains the CP for protein, a second line will develop. 00:03:47.419 --> 00:03:58.769 [upbeat electronic music] 00:03:59.689 --> 00:04:00.409 [music fades] 00:04:00.410 --> 00:04:04.345 Note: If you grow the plants out, you can also use the leaf tissue as well 00:04:04.345 --> 00:04:05.445 instead of the seeds. 00:04:05.812 --> 00:04:10.617 Just take a micro tube, sandwich it between the cap and the leaf. 00:04:12.485 --> 00:04:13.920 Punch a small hole. 00:04:13.920 --> 00:04:16.923 Add your PBS buffer or your distilled water. 00:04:17.123 --> 00:04:19.515 Use this as a mortar and pestle, crush it up, 00:04:19.580 --> 00:04:21.794 and do the lateral flow strip test. 00:04:22.161 --> 00:04:25.164 Is there a difference between these two varieties of soybean? 00:04:25.164 --> 00:04:27.477 Guide a class discussion for students to conclude 00:04:27.477 --> 00:04:30.670 that these two varieties of soybean are fundamentally the same. 00:04:30.903 --> 00:04:32.338 They are both soybeans. 00:04:32.338 --> 00:04:35.341 However, one variety has a protein that the other does not. 00:04:35.541 --> 00:04:38.611 Ask students, "How did the seed obtain that protein?" 00:04:38.945 --> 00:04:40.113 Introduce the concept 00:04:40.113 --> 00:04:43.516 that the genome of a plant can be changed using a variety of methods 00:04:43.950 --> 00:04:45.752 In the following lesson, students will learn 00:04:45.752 --> 00:04:47.993 how these two seed varieties became different 00:04:47.993 --> 00:04:50.375 and explore the impact that a single gene can have. 00:04:50.375 --> 00:04:54.625 [Outro electronic music...]