QuickStixTM Strip Kit EnviroLogix Inc. Corn Seed Teacher's Instructions
QuickStixTM Strip Kit EnviroLogix Inc. Corn Seed Revised 3/03 Teacher’s Instructions Material (per class) 3 containers labeled A, B, and C that students can easily reach into. 400 ml beakers work well. * 3 different varieties of corn seeds, one that expresses the Bt protein**, one that does not express the Bt protein, and one that is an unknown seed variety. Material for each group of four: * 1 QuickStixTM Strip (green colored strip) * 1 disposable 1.5 ml tube * 1 seed from container A, B or C 0.5 ml of tap water * 1 disposable pipette * Material furnished by the Office of Biotechnology to Iowa educators. ** MON810 corn seed courtesy of Garst Seed Company, Slater, IA. Whole seeds should not be placed in public or private landfills. Any whole seeds should be ground up before disposal. How the Test Works Corn that has been genetically modified with a Bt gene will express Bt endotoxin in the seed. The QuickStix kit detects the endotoxins produced by the crystalline proteins Cry1Ab and Cry1Ac. To conduct the test, a seed sample is placed in a small ziplock plastic bag and crushed with a pliers. Then the endotoxins that may be present in the seed are solubilized in water where they can be detected by a lateral flow immunochromatographic test. This assay uses antibodies (on the strips) to detect the presence of Bt protein in the seed. A control line will appear on each strip. If Bt protein is present in the seed, the reaction shows a color change on the strip producing a second line. Each EnviroLogix QuickStix test strip has an absorbent pad at each end. The protective tape with the arrow indicates the end of the strip to insert into the extraction tube. The water with the dissolved endotoxins (if present) will travel up the membrane strip and be absorbed in the larger pad at the top of the strip. The portion of the strip between the protective tape and the absorbent pad at the top of the strip is used to view the reactions as described in the “Interpreting the Results” section of these instructions.
Pre-Lab The instructor should set out three containers labeled A, B, and C. One container will have corn seeds that express the Bt protein. A second container will have corn seeds that do not express the Bt protein. A third container will have corn seeds of an unknown variety. Several groups should test identical containers of seeds to confirm results. Have the students observe the seeds and then record a hypothesis about which seeds they believe to be genetically modified. NOTE: Be very careful to prevent sample-to-sample cross-contamination of seeds, fluids, or disposables. Repeat the protocol for each sample to be tested, using a new tube and plastic bag for each.
Student Instructions Extraction Procedure: Corn Seed 1. Place a single corn seed in a small ziplock bag and seal the bag. 2. Crush the seed using a set of pliers. Be careful not to tear the bag open and lose the contents. Source: EnviroLogix 3. Transfer the crushed seed to a 1.5 ml sample tube. Source: EnviroLogix
4. Using a small transfer pipette, add 0.5 ml of tap water to the sample tube. Source: EnviroLogix 5. Securely cap the sample tube and shake the tube vigorously for 20-30 seconds to extract the protein. Source: EnviroLogix 6. Allow the solid material to settle in the tube 1-2 minutes before placing the QuickStixTM strip into the extract. Source: EnviroLogix
7. Place the QuickStix strip into the extraction solution, being sure to insert the end indicated by the arrows on the protective tape. 8. After inserting the strip into the extraction tube, you will notice liquid traveling up the membrane strip toward the absorbent pad at the top of the strip. Interpreting the Results 9. Soon after the membrane strip is completely wet, a control line will appear on the membrane strip just below the top absorbent pad. This line indicates the strip is functioning correctly. 10. Allow the strip to develop for 10 minutes before making a final interpretation of the results. Record your results on the Class Data Table as instructed by your teacher. 11. The development of a second line below the control line is a positive result that indicates that Bt endotoxin (protein) is present in the seed. A positive result means that the seed was taken from Bt corn. The absence of the second line indicates a negative result for the Bt endotoxin, meaning the seed sample was taken from non-Bt corn. Positive Bt Result Negative Bt Result QuickStixTM is a trademark of Envirologix Inc.
Educators outside of Iowa can get information by mail or by phone from : EnviroLogix 500 Riverside Industrial Parkway Portland, ME 04103-1418 USA (207) 797-0300 Toll free 866-408-4597 FAX (207) 797-7533 Email: info@envirologix.com www.envirologix.com Syngenta Seeds www.syngenta.com
Class Data Table Record the letter of the seed you tested and the result of the test on the chart below. Indicate the result(s) by placing an X in the positive or negative column. Seed Letter Positive (2 lines) Negative (1 line)
Pre-Lab The instructor should set out three containers labeled A, B, and C. One container will have corn seeds that express the Bt protein. A second container will have corn seeds that do not express the Bt protein. A third container will have corn seeds of an unknown variety. Several groups should test identical containers of seeds to confirm .
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background strip each to create two ÒAÓ strip sets. Pair one Color 1 strip and one Color 3 strip with a background strip each to create two ÒBÓ strip sets. Step 2 - Make A and B strip sets. Position each Color strip 3.Ó down from the top of the background strip (the same distance as the width of the background strip).
What are all bfg options? bfg 1.13.0 Usage: bfg [options] [ repo ]-b, --strip-blobs-bigger-than size strip blobs bigger than X (eg '128K', '1M', etc)-B, --strip-biggest-blobs NUM strip the top NUM biggest blobs-bi, --strip-blobs-with-ids blob-ids-file strip blobs with the specified Git object ids-D, --delete-files glob
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Figure 3 Preparing a strip of tape by making a non-stick handle Activity 1: Examining a “T” strip Stick a 15-cm strip of tape on the lab table, sticky side down. This tape forms a standard base for making a “T” (for “Top”) strip. Stick another strip of
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