Protein Expression and Purification Series - Bio-Rad

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Protein Expression and Purification Series Timelines for the Laboratory Course The timeline will depend greatly on the level of the students, which purification method is used, the length of class periods, and whether other techniques and analyses are performed in addition to the basic protocol. To assist in planning for the laboratory course, the following pages provide a workflow, timeline, and helpful hints guide for each purification process. The centrifugation purification process begins below (pg 18–24) with the chromatography instrumentation purification process following (pg 25–32). Centrifugation Purification Workflow 18 Timeline for the Laboratory Course
Protein Expression and Purification Series Centrifugation Purification Laboratory Timeline Note: Tasks that are shaded in grey are preparatory tasks for later stages and may be conducted when spare time is available. Lab Session Timeline for the Laboratory Course: Centrifugation Purification Centrifugation Purification Chapter Task Estimated Duration 0 3A: Advanced Preparation for Centrifugation Purifiation Protocols 1 4: Culturing, Expression, Lysis and SDS-PAGE Analysis for 11 ml Cultures and Purification Protocol for Centrifugation Process 2 4: Culturing, Expression, Lysis and SDS-PAGE Analysis for 11 ml Cultures and Purification Protocol for Centrifugation Process 3 4: Culturing, Expression, Lysis and SDS-PAGE Analysis for 11 ml Cultures and Purification Protocol for Centrifugation Process 19 Module Containing Materials Pour LB/amp plates 30 min Growth and Expression Prepare LB and LB/amp broth 30 min Growth and Expression Prepare IPTG 5 min Growth and Expression Plating E. coli 15 min Growth and Expression Grow E. coli plates overnight at 37°C 16+ hr* Growth and Expression Prepare overnight culture 15 min N/A Grow overnight culture at 37°C 12+ hr* N/A Measure OD600 of overnight culture Prepare uninduced SDS-PAGE samples 30–45 min N/A 15 min SDS-PAGE Electrophoresis Prepare subcultures 15–30 min Growth and Expression Grow subculture 1 hr N/A Measure OD600 of subculture and induce with IPTG 30–45 min Growth and Expression Culture induced cells at 37°C 4–24 hr* N/A *Time indicated for these tasks is not hands-on time. It is the time needed for reactions to run, bacteria to grow, or time to incubate.
Page 1: Biotechnology Explorer TM Protein E
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Page 34 and 35: CHAPTER 1 BACKGROUND Protein Expres
Page 50 and 51: CHAPTER 2 FOCUS QUESTIONS Protein E
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CHAPTER 3A ADVANCE PREP CENTRIFUGAT
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CHAPTER 3A ADVANCE PREP CENTRIFUGAT
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CHAPTER 3B ADVANCE PREP INSTRUMENTA
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CHAPTER 4 11 ml CULTURE PROTOCOL Pr
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CHAPTER 5 100 ml CULTURE PROTOCOL P
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CHAPTER 6 HANDPACKING COLUMNs Prote
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CHAPTER 7 BIOLOGIC LP SYSTEM PROTOC
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CHAPTER 8 BIOLOGIC DUOFLOW PROTOCOL
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CHAPTER 9 DHFR ENZYMATIC ASSAY Prot
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APPENDIX A TIPS AND HELPFUL HINTS P
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APPENDIX A TIPS AND HELPFUL HINTS P
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APPENDIX B RESULTS ANALYSIS Protein
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APPENDIX B RESULTS ANALYSIS Protein
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APPENDIX C RCF TO RPM CONVERSTION P
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APPENDIX D SETTING UP THE SMARTSPEC
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APPENDIX D SETTING UP THE SMARTSPEC
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APPENDIX E MINI-PROTEAN GEL BOX SET
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APPENDIX F BIOLOGIC LP SYSTEM SETUP
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APPENDIX F BIOLOGIC LP SYSTEM SETUP
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APPENDIX G BIOMANUFACTURING Protein
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APPENDIX G BIOMANUFACTURING Protein
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APPENDIX H FOCUS QUESTIONS INSTRUCT
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APPENDIX I GLOSSARY Protein Express
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APPENDIX I GLOSSARY Protein Express
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APPENDIX J REFERENCES, LEGAL NOTES
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Life Science Group Bulletin 1665067
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Protein Expression and Purification Series - Bio-Rad

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