Sci-On® Biology The Biotechnology Education Company ® sed Revi nd a ated Upd EDVO-Kit # S-75 Do Onions, Strawberries and Bananas Have DNA? Storage: See Page 3 for specific storage instructions Experiment Objective: In this experiment, students will learn about the physical nature of DNA by isolating DNA from onions and using the common procedure of DNA spooling. Includes Lesson Plan with Connections to National Content and Skill Standards EDVOTEK, Inc. • 1-800-EDVOTEK • www.edvotek.com EVT 003076K EDVO-Kit # S-75 Do Onions, Strawberries and Bananas Have DNA? Sci-On® Biology Table of Contents Page Experiment Components 3 Experiment Requirements 3 Background Information 5 Experiment Procedures Experiment Overview and General Instructions 7 Activity One - Constructing DNA Models Activity Two - Teacher Demonstration of DNA Spooling 10 Activity Three - Extraction of DNA from Onion Tissue 11 Activity Four - Extraction of DNA from Banana or Strawberry 12 Critical Thinking and Hypothesis Development 13 Study Questions 14 9 Instructor's Guidelines 15 Notes to the Instructor 16 Suggestions for Lesson Plan Content 17 Connectiions to National Standards 18 Pre-Lab Preparations 19 Study Questions and Answers 20 Material Safety Data Sheets All components are intended for educational research only. They are not to be used for diagnostic or drug purposes, nor administered to or consumed by humans or animals. THIS EXPERIMENT DOES NOT CONTAIN HUMAN DNA. None of the experiment components are derived from human sources. EDVOTEK, The Biotechnology Education Company, Sci-On, and InstaStain are registered trademarks of EDVOTEK, Inc.. EVT 003076K EDVOTEK - The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com 21 Sci-On® Biology Do Onions, Strawberries and Bananas Have DNA? EDVO-Kit # S-75 Experiment Components Store entire experiment in the refrigerator This experiment is designed for 10 groups. • • • • • • • Contents DNA Extraction Buffer DNA sample in a capped test tube Transfer pipets Colored Beads (4 colors) Graduated test tubes Spooling rods Salt packet None of the experiment components have been prepared from human sources. Requirements • • • • • • • Fresh onion pieces (scallions, also called green onions, are highly recommended) Ripened fresh strawberry or banana pieces 20 ml beaker Test tubes (13 x 100 mm) 70% clear isopropyl alcohol (rubbing alcohol) Distilled water Ice EDVOTEK - The Biotechnology Education Company® 1-800-EDVOTEK • www.edvotek.com FAX: (301) 340-0582 • email: edvotek@aol.com EVT 003076K EDVO-Kit # Do Onions, Strawberries and Bananas Have DNA? S-75 Sci-On® Biology Online Ordering now available E O DV -TE C H S E RV I C E Technical Service Department 1-800-EDVOTEK (1-800-338-6835) ET Mo Visit our web site for information about EDVOTEK’s complete line of “hands-on” experiments for biotechnology and biology education. Mon - Fri 9:00 am to 6:00 pm ET FAX: (301) 340-0582 Web: www.edvotek.com email: edvotek@aol.com m 6p n - Fri 9 am Please have the following information ready: • Experiment number and title • Kit lot number on box or tube • Literature version number (in lower right corner) • Approximate purchase date EVT 003076K EDVOTEK - The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com Sci-On® Biology EDVO-Kit # Do Onions, Strawberries and Bananas Have DNA? S-75 DNA - Deoxyribonuclease Since the 1800s, it has been known that all living organisms are composed of cells. Organisms such as bacteria are single cells, while very complex organisms, such as humans, are composed of billions of many different cells. 5' DNA plays an important role in two processes. During the process of replication, DNA provides information to copy itself, so genetic information can be passed on from generation to generation of cells. In its second important role, DNA provides instructions for making proteins, which are vital to the maintenance and function of cells. DNA provides information for the order of amino acids required for making various proteins. T A C G 3' C G T A C G The structure of the DNA molecule was determined by James Watson T and Francis Crick in 1953. They A determined that DNA was a double helix consisting of two strands. The Watson and Crick model is often deC G scribed as a spiral ladder. The two strands of DNA are the backbone of 3' 5' the ladder, made of sugar phosphoFigure 1 diester groups. The sugar backbone acts as a support for the rungs of the ladder. These rungs are composed of the chemical bases Adenine, Guanine, Cytosine, and Thymine. The first letters of these bases, A,G,C, and T, are used by scientists to designate the order of the bases within the DNA strands. Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com Background Information In 1868, a Swiss biologist named Friedrich Meischer carried out research which indicated that the nucleus of cells contains a material which he called nucleic acid. But, it wasn’t until much later in the 1940’s that the nucleic acid, deoxyribonucleic acid (DNA), was recognized as the carrier of genetic information. EDVO-Kit # S-75 Do Onions, Strawberries and Bananas Have DNA? Sci-On® Biology DNA - Deoxyribonuclease Background Information The bases are always arranged in pairs. When A occurs on one strand, T occurs on the opposite strand. Similarly, G and C are opposite DNA strands. The bases are held together by weak bonds which are shown as dashed lines in Figure 1. 1. 2. 3. DNA can be extracted from the nucleus of cells by adding an aqueous buffered extraction solution to cells. The cells are chemically lysed (broken open) and DNA from chromosomes is released. This procedure is known as cell lysis. DNA is soluble in water and cannot be seen, but it is insoluble in alcohol. Purification procedures for nucleic acids usually include precipitation with alcohol in the presence of salt. Since rubbing alcohol (isopropyl alcohol) has a lower density than water, it will form a second layer above the DNA solution. A spooling rod is used to spool the two liquids at the interface of the two phases to separate the DNA from 1 the solution. The DNA will appear as a viscous, clotted mass as it is collected on the spooling rod (Figure The DNA is inside the cell. 2). The amount of DNA spooled is a The cell wall is disrupted. consequence of the size of the DNA The DNA is spooled onto a rod. fragments which are much larger than 2 the small bio-molecules such as amino acids and small carbohydrate sugars. 3 Figure 2 In this experiment, DNA will be isolated in one of two ways: 1) DNA will be spooled from a buffer solution containing salts; 2) DNA will be extracted from tissues such as onion, banana, or strawberry and spooled from solution. DNA will be observed by the naked eye on the spooling rod. Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com Sci-On® Biology Do Onions, Strawberries and Bananas Have DNA? EDVO-Kit # S-75 Experiment Overview and General Instructions Experiment Content Objective Working Hypothesis DNA can be extracted from the cells, and isolated from solution by spooling. LABORATORY SAFETY 1. Gloves and goggles should be worn routinely as good laboratory practice. 2. Exercise extreme caution when working with equipment that is used in conjunction with the heating and/or melting of reagents. 3. Do not mouth pipet reagents - use pipet pumps. 4. Exercise caution when using any electrical equipment in the laboratory. 5. Always wash hands thoroughly with soap and water after handling reagents or biological materials in the laboratory. Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com The Experiment The objectives of this experiment are to learn: • how to isolate DNA from onion, banana or strawberry cells • what DNA looks like in solution • how to spool DNA from solution • what DNA looks like after it is spooled from solution • the structure of DNA from building a simulated DNA model • the function of DNA EDVO-Kit # S-75 Do Onions, Strawberries and Bananas Have DNA? Sci-On® Biology Experiment Overview and General Instructions before you start the experiment The Experiment Read all instructions before starting the experiment. LAB NOTEBOOK Predict the outcomes from the experiment and the control. Record your predictions in your lab notebook or on a separate sheet of paper. Before starting the Experiment: • • Write a hypothesis that reflects the experiment. Predict experimental outcomes. During the Experiment: • Record (draw) your observations, or photograph the results. Following the Experiment: • Formulate an explanation from the results. • Determine what could be changed in the experiment if the experiment were repeated. • Write a hypothesis that would reflect this change. Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com Sci-On® Biology Do Onions, Strawberries and Bananas Have DNA? EDVO-Kit # S-75 Activity One - Constructing DNA Models 1. Each group of students will get a set of beads of various colors. 2. The teacher will assign a base designation for four of the colors. For example: Red = Adenine (A) Blue = Green =Guanine (G) Yellow = Thymine (T) Cytosine (C) A-T base pair G-C base pair How many A-T or G-C base pairs were you able to put together? Putting All the Student Base Pair Models Together: 4. With the help of your teacher, and the rest of the class, build a model of the DNA ladder. Attach the ends of several bead sections together to form two long strands of double-stranded DNA. 7. In your own words, describe the structure of DNA Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com The Experiment 3. Sort all the beads by color. Based upon the beads you received, correctly put the colored beads together in two separate strands to form one or more of the following base pair models: 10 EDVO-Kit # S-75 Do Onions, Strawberries and Bananas Have DNA? Sci-On® Biology Activity Two - Teacher Demonstration of DNA Spooling The Experiment In this activity, DNA will be spooled from an aqueous buffered solution by adding salt and overlaying with isopropyl alcohol. The isopropyl alcohol (clear rubbing alcohol) should be ice cold (stored in a freezer at least 2 hours before use). Just before the demonstration, place the DNA solution and the alcohol on ice. FREEZER The Experiment: It is important that the 70% clear isopropyl alcohol is very cold before beginning this demonstration. - 1. Add two pinches of salt to a small beaker or clear graduated test tube. 2.Use a transfer pipet to add 2 ml of distilled water and swirl to dissolve the salt. 3.Use a transfer pipet to transfer all of the DNA solution to the salt water. Stir or swirl to mix. The total volume should be approximately 3 ml. 4.Using a transfer pipet, carefully overlay, the DNA solution with two volumes of very cold 70% clear isopropyl alcohol (about 6 ml). Let the cold alcohol gently flow down the side of the beaker or test tube. Do not mix the two solutions. 5. Place the end of the spooling rod just below the line separating the two solutions (the interface). Twirl the rod with two fingers and spool the DNA from the solution. 6. Observe the DNA and record your observations. The Control 7. Add two pinches of salt to a small beaker or clear graduated test tube. 8. Add distilled water up to the 2 ml level of the tube and swirl to dissolve the salt. 9. Add another 1 ml of distilled water (instead of DNA). Swirl to mix. 10. Layer the cold alcohol on top of the water solution in the same manner as step #4 of the experiment. Do not mix the two solutions. 11. Place the end of the spooling rod just below the line separating the two solutions (the interface). Twirl the rod with two fingers and spool. 12. Record your observations for the control . Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com Sci-On® Biology Do Onions, Strawberries and Bananas Have DNA? EDVO-Kit # 11 S-75 Activity Three - Extraction of DNA from Onion Tissue In this activity, DNA is extracted from onion tissue using an extraction buffer and spooled from solution by overlaying the liquid with ice cold isopropyl alcohol. The Experiment 2.Using a transfer pipet, add DNA Extraction Buffer until it reaches the 3 ml level mark on the tube. Mince the onion cube with the tip of a spooling rod, pencil or pen (mash the onion very well). This will release the cellular contents and the DNA in the onion cells. 3. With a transfer pipet, carefully remove 2 ml of the liquid layer from the test tube and transfer it into a second, clean test tube. Try to minimize carry-over of the onion tissue. 4. Carefully overlay the liquid with 4 ml of very cold 70% clear Isopropyl alcohol. 5. Place a spooling rod into the test tube and twirl it with two of your fingers. Gently rotate the rod at the interface of the two phases. The DNA will begin to spool (wrap) around the rod. Gently lift the rod out of the solution from time to time and observe the DNA substance attached to it. 6. After spooling for several minutes, remove the rod from the test tube to observe the DNA. The DNA will appear as a viscous material adhering to the splint (it may be grayish in color if a pencil was used to mince the onion tissue). LAB NOTEBOOK 7. Record your observations of the onion DNA. The CONTROL 8. Based upon what you have previously learned, outline and perform the control for this activity. Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com The Experiment 1. Carefully slice a 5 x 5 x 5 mm cube (about the size of a pencil eraser) from the main body of the onion (not the root tip), and place it in one of the test tubes provided. 12 EDVO-Kit # S-75 Do Onions, Strawberries and Bananas Have DNA? Sci-On® Biology Activity Four - Extraction of DNA from Strawberries or Banana In this activity, DNA is extracted from a banana or strawberries in the same manner as the extraction of DNA from onion. The Experiment 1. Carefully slice a 5 x 5 x 5 mm cube from the fleshy body of the banana or strawberry, and place it in a clean test tube. 2.Using a transfer pipet, add DNA Extraction Buffer until it reaches the 3 ml mark on the tube. Mince the cube with the tip of a spooling rod, pencil or pen to release the cellular contents. 3. With a transfer pipet, carefully remove 2 ml of the liquid layer from the test tube and transfer it into a second, clean test tube. Try to minimize carry-over of the banana or strawberry tissue. 4. Carefully overlay the liquid with 4 ml of very cold 70% clear Isopropyl alcohol. 5. Place a spooling rod into the test tube and twirl it with two of your fingers. Gently rotate the rod at the interface of the two phases. The DNA will begin to spool (wrap) around the rod. Gently lift the rod out of the solution from time to time and observe the DNA substance attached to it. 6. After spooling for several minutes, remove the rod from the test tube to observe the DNA. The DNA will appear as a viscous material adhering to the splint (it may be grayish in color if a pencil was used to mince the onion tissue). LAB NOTEBOOK 7. Record your observations of the banana or strawberry DNA. Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com Sci-On® Biology Do Onions, Strawberries and Bananas Have DNA? EDVO-Kit # 13 S-75 Critical Thinking and Hypothesis Development LAB NOTEBOOK Record the following in your Laboratory Notebook or on a separate sheet of paper: 1. What are the variables in this experiment? 3. Write a hypothesis that would reflect these changes. Study Questions LAB NOTEBOOK Record the answers to the following Study Questions in your Laboratory Notebook or as instructed by your teacher. 1. Describe the appearance of the isolated DNA. 2. What is a nucleus? 3. What are nucleotides? 4. What are chromosomes? 5. What does cell lysis mean? 6. Why did the alcohol layer on top of the DNA solution? 7. Why was it important not to mix the alcohol and DNA solutions? 8. What properties of DNA allow spooling? 9. What difficulties did you have in spooling? Why? 10. How many chromosomes do humans have? Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com The Experiment 2. What would you have changed in the experiment if you had to do it over again? Why? Predict outcomes. 14 EDVO-Kit # S-75 Do Onions, Strawberries and Bananas Have DNA? Sci-On® Biology EVT 003076K EDVOTEK - The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com Sci-On® Biology Do Onions, Strawberries and Bananas Have DNA? EDVO-Kit # S-75 Instructor’s Guide Class size, length of laboratory sessions, and availability of equipment are factors which must be considered in the planning and the implementation of this experiment with your students. These guidelines can be adapted to fit your specific set of circumstances. If you do not find the answers to your questions in this section, a variety of resources are continuously being added to the EDVOTEK web site. In addition, Technical Service is available from 9:00 am to 6:00 pm, Eastern time zone. Call for help from our knowledgeable technical staff at 1-800-EDVOTEK (1-800-338-6835). E O DV -TE C H S E RV I C E Technical Service Department 1-800-EDVOTEK ET (1-800-338-6835) Mo Mon - Fri 9:00 am to 6:00 pm ET FAX: (301) 340-0582 Web: www.edvotek.com email: edvotek@aol.com m 6p n - Fri 9 am Please have the following information ready: • Experiment number and title • Kit lot number on box or tube • Literature version number (in lower right corner) • Approximate purchase date Online Ordering now available Visit our web site for information about EDVOTEK’s complete line of “hands-on” experiments for biotechnology and biology education. EDVOTEK - The Biotechnology Education Company® 1-800-EDVOTEK • www.edvotek.com FAX: (301) 340-0582 • email: edvotek@aol.com EVT 003076K 15 16 EDVO-Kit # S-75 Do Onions, Strawberries and Bananas Have DNA? Sci-On® Biology Notes to the Instructor: Instructor’s Guide EXPERIMENT Hints and Help EDVOTEK experiments are easy to perform and are designed for maximum success in the classroom setting. However, even the most experienced students and teachers occasionally encounter experimental problems or difficulties. The EDVOTEK web site provides a variety of resources which are continuously being updated and added. If you do not find the answers to your questions in this section or at the EDVOTEK web site, Technical Service is available from 9:00 am to 6:00 pm, Eastern time zone. Call for help from our knowledgeable technical staff at 1-800-EDVOTEK (1-800-338-6835). Hints for Maximizing Success 1. Isopropyl alcohol should be placed in the freezer to ensure that it is very cold. Keep the alcohol on ice while students are using it to overlay their DNA solution for spooling. 2. DNA spooling results from onions, strawberries or banana will vary, but it is recommended that they be as fresh as possible. The material spooled from bananas will contain a mixture of DNA and carbohydrates. Clean-up and disposal of materials The DNA solutions and alcohol can be disposed down the drain. All other materials can be disposed in regular solid waste (trash). Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com Sci-On® Biology Do Onions, Strawberries and Bananas Have DNA? EDVO-Kit # 17 S-75 Suggestions for Lesson Plan Content 1. Conduct a discussion of DNA, the material which controls who we are and what we look like. 2. Give students an overview of the structure of DNA. 3. Write the following words on the board for discussion: Nucleus Chromosome Gene DNA Double helix Nucleotide (base) Deoxyribose Phosphodiester bond Base pair Adenine Guanine Cytosine Thymine Use pictures or drawings from textbooks and other resources to help explain the vocabulary words to the students. 4. Focus on the following concepts to ensure student understanding, to maximize the benefits of the “hands-on” experience. • • • • • • • • DNA is found in the cell nucleus of human cells and in the nuclear area in bacteria. DNA is made up of two strands. Each strand has a sequence of nucleotides (bases) placed in a specific order. The bases (nucleotides) are called Adenine (A), Guanine (G), Thymine (T), and Cytosine (C). The two strands are held together because the bases form physical bonds (known as hydrogen bonds) with each other in very specific ways. Adenine base-pairs with Thymine and Cytosine base-pairs with Guanine. The strands are joined to form a double helix. A chromosome contains the coding segments called genes. Each gene codes for a specific RNA which provides the required information to make proteins. 5. List and discuss with students the essential parts of an experiment. • • • • • • • • writing a logical hypothesis making careful observations differentiating between an experiment and an control dentifying variables predicting experimental outcomes recording results in a concise and accurate manner drawing valid interpretations of results formulating alternative explanations Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com Instructor’s Guide This teacher-generated lesson plan outline can be used as a guideline for classroom discussion. Connections to the National Content and Skills Standards follow the plan. 18 EDVO-Kit # S-75 Do Onions, Strawberries and Bananas Have DNA? Sci-On® Biology Connections to National Standards Content Standards 1. Students will develop abilities necessary to do scientific inquiry. Instructor’s Guide • Questions will be answered through scientific investigation. 2. Students will develop an understanding through inquiry. Students will: • • • • • • • • develop a logical hypothesis. make careful observations. interpret results correctly. understand differences between the experiment and the control. identify and control variables. predict experimental outcomes. formulate explanations from evidence. recognize and analyze alternative explanations. 3. Students will use equipment, materials, and procedures for experimentation and direct investigation of phenomena. • Students will understand DNA isolation by spooling. 4. Students will develop an understanding of structure and function of the molecule of heredity - DNA. Students will: • understand nucleotide composition and its role as the building block. • develop an understanding of the physical nature of DNA. Skill Standards In this experiment students will learn to spool and stain chromosomal DNA. Analysis of the experiment will provide students the means to transform an abstract concept into a concrete experience. Students will be able to: 1. Isolate DNA from a suspended solution by alcohol precipitation and spooling. 2. Use scientific equipment such as calibrated pipets and graduated cylinders to make metric based measurement. 3. Make careful observations and record results. 4. Predict experimental outcomes. 5. Share and compare results with other students. 6. Develop alternate options for DNA spooling, make predictions and compare with others students. 7. Explain the implication of DNA extraction for use as materials for DNA experiments. Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com Sci-On® Biology EDVO-Kit # Do Onions, Strawberries and Bananas Have DNA? 19 S-75 Pre-Lab Preparations On the day of the lab: 1. Thoroughly chill the 70% isopropyl alcohol (rubbing alcohol) before the students perform the experiment. • Store it in the freezer for several hours or overnight to ensure that it is very cold. •Using a calibrated transfer pipet, transfer 6 ml of 70% isopropyl alcohol into 20 test tubes. • FREEZER Place the alcohol-filled tubes back in the freezer or on ice until students are ready to use the alcohol. 2. Each student group requires: • • • • • • Sets of beads (various colors) Graduated test tube Spooling rod Transfer pipet DNA extraction buffer - 3-4 ml Cold isopropyl alcohol - 4 ml Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1996, 1997, 1998, 1999, 2003, 2006, EDVOTEK, Inc., all rights reserved EVT 003066K The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com Instructor’s Guide Please refer to the kit insert for the Answers to Study Questions EDVOTEK Section V - Reactivity Data Material Safety Data Sheet IDENTITY (As Used on Label and List) Emergency Telephone Number Manufacturer's Name EDVOTEK, Inc. Telephone Number for information Address (Number, Street, City, State, Zip Code) Date Prepared 14676 Rothgeb Drive Rockville, MD 20850 Conditions to Avoid X Stable Incompatibility Note: Blank spaces are not permitted. If any item is not applicable, or no information is available, the space must be marked to indicate that. Concentrated Chromosomal DNA Section I Unstable Stability May be used to comply with OSHA's Hazard Communication Standard. 29 CFR 1910.1200 Standard must be consulted for specific requirements. ® (301) 251-5990 (301) 251-5990 01/15/06 Hazardous Decomposition or Byproducts Toxic fumes of: Carbon Monoxide, Carbon dioxide, nitrogen oxides Hazardous Polymerization Conditions to Avoid May Occur X Will Not Occur None Section VI - Health Hazard Data Route(s) of Entry: Inhalation? Skin? Yes Health Hazards (Acute and Chronic) Yes Ingestion? Yes Toxicological properties have not been thoroughly investigated Carcinogenicity: IARC Monographs? NTP? No data Signature of Preparer (optional) None Iron, steel, alumium, or copper No data OSHA Regulation? No data No data Signs and Symptoms of Exposure Can cause coughing, wheezing, respiratory distress, and diarrhea. Skin/eye irritation. Section II - Hazardous Ingredients/Identify Information Hazardous Components [Specific Chemical Identity; Common Name(s)] OSHA PEL N-Lauroylsarcosine sodium Sarcosine, N-dodecyl-, sodium salt CAS # 7631-98-3 Section III - Physical/Chemical Characteristics Other Limits Recommended ACGIH TLV % (Optional) Treat supportively and symptomatically No data Specific Gravity (H 0 = 1) 2 No data Vapor Pressure (mm Hg.) No data Melting Point No data Evaporation Rate (Butyl Acetate = 1) No data No data Solubility in Water No data Emergency First Aid Procedures Section VII - Precautions for Safe Handling and Use Boiling Point Vapor Density (AIR = 1) Medical Conditions Generally Aggravated by Exposure Steps to be Taken in case Material is Released for Spilled Mop up with absorbent material. Dispose of properly. Waste Disposal Method Bury in landfill site approved for disposal of chemical and hazardous waste. Observe all federal, state, and local laws. Precautions to be Taken in Handling and Storing Avoid contact. Do not store in contact with iron, steel, aluminum, or copper Soluble Appearance and Odor Other Precautions None Clear liquid, no odor Section IV - Physical/Chemical Characteristics Flash Point (Method Used) LEL Flammable Limits No data UEL No data No data Extinguishing Media Section VIII - Control Measures Respiratory Protection (Specify Type) Ventilation Carbon dioxide, dry chemical powder, alcohol or polymer foam, water spray Special Fire Fighting Procedures Protective Gloves NIOSH/MSHA approved respirator Local Exhaust No Special Mechanical (General) No Other Wear SCBA and prot ective clothing to prevent contact w/ skin & eyes Other Protective Clothing or Equipment None Work/Hygienic Practices Unusual Fire and Explosion Hazards IDENTITY (As Used on Label and List) Emergency Telephone Number Manufacturer's Name EDVOTEK, Inc. Address (Number, Street, City, State, Zip Code) Telephone Number for information Date Prepared 14676 Rothgeb Drive Rockville, MD 20850 Do not ingest. Avoid contact with skin, eyes, and clothing. Conditions to Avoid X Stable Incompatibility Note: Blank spaces are not permitted. If any item is not applicable, or no information is available, the space must be marked to indicate that. Concentrated Buffer Section I Rubber boots Unstable Stability May be used to comply with OSHA's Hazard Communication Standard. 29 CFR 1910.1200 Standard must be consulted for specific requirements. ® None Chem. splash proof Section V - Reactivity Data Material Safety Data Sheet EDVOTEK Eye Protection Rubber/chemical resistant gloves None (301) 251-5990 (301) 251-5990 Excessive heat, sparks or open flame Acids, Aluminum, metals, oxidizers (strong) Hazardous Decomposition or Byproducts Thermal decomposition products of toxic and hazardous oxides of Carbon, Nitrogen, and Sodium. Hazardous Polymerization Conditions to Avoid May Occur X Will Not Occur None Section VI - Health Hazard Data Route(s) of Entry: Inhalation? Skin? Yes Yes Yes Ingestion? Health Hazards (Acute and Chronic) Moderately toxic by ingestion-systemic toxicity may result. May chelate lead, magnesium, zinc, & trace metals-possibly causing increased absorption and increasing total body storage. 01//15/06 Carcinogenicity: NTP? No data Signature of Preparer (optional) No data IARC Monographs? No data OSHA Regulation? No data Signs and Symptoms of Exposure Section II - Hazardous Ingredients/Identify Information Hazardous Components [Specific Chemical Identity; Common Name(s)] OSHA PEL ACGIH TLV Ethylenediaminetetraacetic acid No data No data C10-H14-08-N2.2Na CAS #139-33-3 Section III - Physical/Chemical Characteristics Boiling Point No data Vapor Pressure (mm Hg.) No data Vapor Density (AIR = 1) Solubility in Water Appearance and Odor No data Other Limits Recommended No data % (Optional) No data Medical Conditions Generally Aggravated by Exposure Renal or heart disease, potassium deficiency, insulin dependent diabetes, seizures or intracranial lesions Emergency First Aid Procedures Treat supportively and symptomatically Section VII - Precautions for Safe Handling and Use No data Specific Gravity (H 0 = 1) 2 Melting Point No data Evaporation Rate (Butyl Acetate = 1) No data Steps to be Taken in case Material is Released for Spilled Mop up with absorbent material. Containerize to dispose of properly. Waste Disposal Method Observe all federal, state, and local laws. Precautions to be Taken in Handling and Storing Store away from strong oxidizers or heat. Avoid skin/eye contact. Soluble Other Precautions None Clear liquid, no odor Section IV - Physical/Chemical Characteristics Flash Point (Method Used) Mucous membrane irritation, eye/skin irritation, irritating to gastrointestinal system. No data Flammable Limits LEL No data UEL No data Extinguishing Media Dry chemical, carbon dioxide, halon, water spray or standard foam Special Fire Fighting Procedures Move container from fire area if possible. Thermal decomposition products may include toxic and hazardous oxides of Carbon, nitrogen and sodium. Unusual Fire and Explosion Hazards Section VIII - Control Measures Respiratory Protection (Specify Type) Chemical cartridge respirator with full facepiece and organic vapor cartridge Special Yes None Local Exhaust Ventilation Protective Gloves Mechanical (General) Gen. dilution vent. sys Other None Yes Other Protective Clothing or Equipment Work/Hygienic Practices Eye Protection Splash-proof goggles Impervious clothing to prevent skin contact Do not ingest. Avoid contact with skin, eyes, and clothing. Section V - Reactivity Data Material Safety Data Sheet EDVOTEK ® IDENTITY (As Used on Label and List) Section I Stability May be used to comply with OSHA's Hazard Communication Standard. 29 CFR 1910.1200 Standard must be consulted for specific requirements. Emergency Telephone Number Manufacturer's Name EDVOTEK, Inc. Address (Number, Street, City, State, Zip Code) Telephone Number for information Date Prepared 14676 Rothgeb Drive Rockville, MD 20850 Conditions to Avoid X Incompatibility Note: Blank spaces are not permitted. If any item is not applicable, or no information is available, the space must be marked to indicate that. Extraction Buffer Unstable Stable (301) 251-5990 (301) 251-5990 Strong oxidizing agents, protect from moisture Carbon monoxide, carbon dioxide, nitrogen oxides, hydrogen bromide gas Hazardous Decomposition or Byproducts Hazardous Polymerization Conditions to Avoid May Occur X Will Not Occur Section VI - Health Hazard Data Route(s) of Entry: Inhalation? Ingestion? Skin? Yes Yes Yes Health Hazards (Acute and Chronic) Harmful if swallowed, inhaled, or absorbed through skin. Material is extremely destructive to tissue of the mucous membranes and upper respiratory tract, eyes and skin. 01/15/06 Carcinogenicity: Signature of Preparer (optional) IARC Monographs? NTP? OSHA Regulation? N/A Signs and Symptoms of Exposure Section II - Hazardous Ingredients/Identify Information Hazardous Components [Specific Chemical Identity; Common Name(s)] OSHA PEL ACGIH TLV Other Limits Recommended % (Optional) 5144-89-8 60-00-4 57-09-0 Section VII - Precautions for Safe Handling and Use Steps to be Taken in case Material is Released for Spilled Boiling Point 100°C Specific Gravity (H 0 = 1) 2 1.020 Vapor Pressure (mm Hg.) No data Melting Point 100°C No data Evaporation Rate (Butyl Acetate = 1) No data Vapor Density (AIR = 1) Appearance and Odor Special Fire Fighting Procedures Waste Disposal Method Dissolve or mix the material w/ combustible solvent and burn in a chemical incinerator equipped with an afterburner. Ventilate area and wash spill site after material pick up is complete Precautions to be Taken in Handling and Storing Other Precautions Clear liquid Flash Point (Method Used) Evacuate area. Cover with dry lime or soda ash-pick up. keep in a closed container and hold for waste disposal. Observe all federal, state, and local laws. Yes Section IV - Physical/Chemical Characteristics Extinguishing Media Emergency First Aid Procedures Immediately flush eyes or skin w/ copious amounts of water for at least 15 minutes while removing Section III - Physical/Chemical Characteristics Solubility in Water Medical Conditions Generally Aggravated by Exposure Flammable Limits Water spray Wear SCBA and protective clothing to prevent contact w/ skin and eyes. Unusual Fire and Explosion Hazards Emits toxic fumes under fire conditions Use only in a chemical fume hood. LEL UEL Section VIII - Control Measures Respiratory Protection (Specify Type) Ventilation Wear appropriate NIOSH/MSHA aprroved respirator. Special Local Exhaust Mechanical (General) Protective Gloves safety gloves Other Protective Clothing or Equipment Work/Hygienic Practices Other Eye Protection safety goggles Safety shower and eye bath, face shield Wash thoroughly after handling, keep tightly closed.