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2 EDVO-Kit # 953 xxx Water Quality Testing III: Multiplex PCR Analysis for Water Contaminants Table of Contents Experiment Components 3 Experiment Requirements 4 Background Information 5 Experiment Procedures Experiment Overview and General Instructions 9 Laboratory Safety 10 Module I: Isolation of DNA 11 Module II: PCR Amplification of the Multiplex Water Contaminants 12 Module III: Separation of PCR Reactions by Agarose Gel Electrophoresis 14 Study Questions 16 Instructor's Guidelines Notes to the Instructor 18 Pre-Lab Preparations 21 Experiment Results and Analysis 23 Study Questions and Answers 24 Appendices A PCR Experimental Success Guidelines 26 B Polymerase Chain Reaction Using Three Water baths 28 C Preparation and Handling of PCR Samples With Wax 29 D 1.0% Agarose Gel Preparation 30 E 1.0% Agarose Gels - Quantity Preparations 31 F Staining and Visualization of DNA with InstaStain® Ethidium Bromide Cards 32 G InstaStain® Methylene Blue: One Step Staining and Destaining 33 Material Safety Data Sheets 34 EVT 090205K EDVOTEK, The Biotechnology Education Company, and InstaStain are registered trademarks of EDVOTEK, Inc.. Ready-to-Load and UltraSpec-Agarose are trademarks of EDVOTEK, Inc. EDVOTEK - The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
Water Quality Testing III: Multiplex PCR Analysis for Water Contaminants EDVO-Kit # 953 3 Components & Requirements This experiment is designed for 20 students working in groups of five. Storage A Tubes with PCR reaction pellets Room Temperature Each PCR reaction pellet contains • dNTP Mixture • Taq DNA Polymerase Buffer • Taq DNA Polymerase • MgCl 2 B Primer mixture -20°C Freezer (contains E. coli-specific primers, Bacillus subtilis-specific primers, and Serratia marcescens-specific primers C 200 base pair ladder -20°C Freezer D E. coli DNA for positive control -20°C Freezer E Tris buffer -20°C Freezer F Proteinase K Room temperature G Potassium Acetate Room temperature H DNA Extraction buffer Room temperature • E. coli culture Room temp. desiccated • Bacillus subtilis culture Room temp. desiccated • Serratia marcescens culture Room temp. desiccated Reagents & Supplies (Store all components below at room temperature) 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. • UltraSpec-Agarose • Electrophoresis Buffer (50x) • 10x Gel Loading Solution • InstaStain® Ethidium Bromide • InstaStain® Methylene Blue • Microcentrifuge Tubes • PCR tubes (0.2 ml - for thermal cyclers with 0.2 ml template) • 50 ml Conical tube • Wax beads (for water bath option or thermal cyclers without heated lid) EDVOTEK - The Biotechnology Education Company® 1-800-EDVOTEK • www.edvotek.com FAX: (301) 340-0582 • email: info@edvotek.com EVT 090205K
Page 1: The Biotechnology Education Company
Page 5 and 6: Water Quality Testing III: Multiple
Page 27 and 28: Appendix A PCR Experimental Success
Page 29 and 30: Appendix C Preparation and Handling
Page 31 and 32: Appendix E 1.0% Agarose Gels - Quan
Page 33 and 34: Appendix G Staining and Visualizati
Page 35 and 36: Material Safety Data Sheets Full si

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