1. Recombinant DNA Training for Research Group Members How to Use this Training Template Principal Investigators (PIs) whose research involves recombinant DNA are required by NIH to provide their research group members training on lab-specific hazards and emergency procedures. The Berkeley Biosafety Program(bso@berkeley.edu) has prepared this PowerPoint presentation template to assist PIs in meeting this requirement. Text in italics [example] indicates where the template should be modified to accurately reflect your lab hazards and procedures. bso@berkeley.edu

2. Working with Recombinant DNA in the _[name of PI]_ Laboratory [Name of PI] [Name of Department] [Date Training Is Given]

3. Basic Requirements Research involving recombinant DNA is closely regulated by the National Institutes of Health (NIH), and requires a Biological Use Authorization (BUA) approved by the Berkeley campus Committee on Laboratory & Environmental Biosafety (CLEB). CLEB works with the campus Biosafety Program, which can provide additional guidance (bso@berkeley.edu) bso@berkeley.edu

4. Basic Requirements (Cont’d) All work involving recombinant DNA – regardless of funding source -- must adhere to the mandatory NIH “Guidelines for Research Involving recombinant DNA Molecules” found at http://oba.od.nih.gov/rdna/nih_guidelines _oba.html http://oba.od.nih.gov/rdna/nih_guidelines _oba.html http://oba.od.nih.gov/rdna/nih_guidelines _oba.html

5. Purpose of This Training Section IV-B-7-d-(2) of the NIH Guidelines The NIH requires that the PI provide lab- specific biosafety training on the hazards of the specific experimental procedures used in their laboratory to members of their research group. One way to document this training is to record it in the minutes of a group meeting, listing all participants.

6. Contents of This Training CLEB Administrative Requirements CLEB Administrative Requirements What are the NIH Guidelines? What are the NIH Guidelines? Role of the Investigator/Lab Staff Role of the Investigator/Lab Staff Description of the Experiment-BUA Description of the Experiment-BUA –The Agents Used –The Template Risk Assessment/Biosafety Level Risk Assessment/Biosafety Level –Good Work Practices –Personal Protective Equipment –Engineering Controls Waste Disposal/Decontamination Waste Disposal/Decontamination Emergency Procedures Emergency Procedures

7. CLEB Administrative Requirements CLEB requires the following from every PI approved to use recombinant DNA: CLEB requires the following from every PI approved to use recombinant DNA: –Train all research group members on lab- specific hazards and procedures –Report incidents involving recombinant DNA immediately to EH&S –Submit a BUA amendment for any change in personnel or Scope of Work

8. What are the NIH Guidelines? Key Sections of the NIH Guidelines Appendix B-List of Agents Appendix B-List of Agents Section III-Types of Experiments involving recombinant DNA Section III-Types of Experiments involving recombinant DNA Appendix G-Level of Containment Appendix G-Level of Containment Appendix K-Large Scale Appendix K-Large Scale Appendix M-Gene Transfer/Human Subjects Appendix M-Gene Transfer/Human Subjects Appendix P-Transgenic Plants Appendix P-Transgenic Plants Appendix Q-Transgenic Animals Appendix Q-Transgenic Animals Section IV-Responsibilities of PI, IBC Section IV-Responsibilities of PI, IBC

9. PI RESPONSIBILITIES Section IV-B-7: ROLE OF THE PI On behalf of the institution, the Principal Investigator is responsible for full compliance with the NIH Guidelines for any recombinant DNA research. The PI must: The PI must: Not initiate or modify recombinant DNA research which requires CLEB approval until that research has been approved by CLEB and has met all other requirements of the NIH Guidelines. Not initiate or modify recombinant DNA research which requires CLEB approval until that research has been approved by CLEB and has met all other requirements of the NIH Guidelines. Report any significant problems, violations of the NIH Guidelines, or any significant research-related accidents and illnesses to the Biological Safety Officer and CLEB. Report any significant problems, violations of the NIH Guidelines, or any significant research-related accidents and illnesses to the Biological Safety Officer and CLEB.

10. OTHER PI RESPONSIBILITIES ResponsibilityNIH Guideline Reference Train Lab Staff (Micro technique)Section IV-B-7-a, d Train Lab Staff (Micro technique)Section IV-B-7-a, d Ship Agents SafelySection IV-B-7-a Ship Agents SafelySection IV-B-7-a Follow Emergency PlansSection IV-B-7-a Follow Emergency PlansSection IV-B-7-a Notify NIH of novel host/vectorsSection IV-B-7-b Notify NIH of novel host/vectorsSection IV-B-7-b Update ProtocolSection IV-B-7-c Update ProtocolSection IV-B-7-c Notify IBC/CLEB of incident Section IV-B-7-e Notify IBC/CLEB of incident Section IV-B-7-e Make Protocol AvailableSection IV-B-7-d Make Protocol AvailableSection IV-B-7-d Offer Medical SurveillanceSection IV-B-7-d Offer Medical SurveillanceSection IV-B-7-d Supervise Lab MembersSection IV-B-7-e Supervise Lab MembersSection IV-B-7-e Maintain Safety EquipmentSection IV-B-7-e Maintain Safety EquipmentSection IV-B-7-e

11. Description of the Experiment-BUA The Scope of Work [Cut and paste scope of work narrative from your BUA] [Cut and paste scope of work narrative from your BUA]

12. Description of the Experiment-BUA The Agents Used [List the agents, recombinant DNA host/vector systems, human cell lines, packaging cell lines, and clinical specimens used in the experiment. Include all etiological agents.]

13. Description of the Experiment-BUA The Risks That May Affect Containment Level These are some of the common experimental risks associated with our research: [modify list as appropriate] These are some of the common experimental risks associated with our research: [modify list as appropriate] –Route(s) of exposure –volumes of culture –aerosol generating procedures –Procedures involving sharps –high concentrations –the use of vectors and vulnerable animals –inserted genetic sequences that increase virulence –the use of drug/antibiotic resistant strains

14. Description of the Experiment-BUA The Risks That May Affect Containment Level These are the symptoms of exposure with the agents we use: These are the symptoms of exposure with the agents we use: –[List symptoms of exposure]

15. Our Recombinant DNA Experiment Template [Cut and paste from BUA form:] Agent Characteristics: Agent Characteristics: Routes of Exposure: Routes of Exposure: Host: Host: Vector: Vector: Nature of inserted sequences: Nature of inserted sequences: Source of inserted sequences: Source of inserted sequences: Types of manipulation: Types of manipulation: Attempt to express foreign gene: Attempt to express foreign gene: Protein produced: Protein produced: Containment: Containment: Section of Guidelines: Section of Guidelines:

16. Risk Assessment/Biosafety Level Good Work Practices [Modify list as appropriate] Wash Hands Frequently Wash Hands Frequently Use Personal Protective Equipment and Engineering Controls Use Personal Protective Equipment and Engineering Controls Minimize Use of Sharps Minimize Use of Sharps Do Not Eat, Drink, or Smoke in the lab Do Not Eat, Drink, or Smoke in the lab Disinfect Work Areas Disinfect Work Areas

17. Risk Assessment/Biosafety Level Personal Protective Equipment [Modify list as appropriate] The following personal protective equipment must be worn when handling agents in the tissue culture room and the biosafety cabinet: The following personal protective equipment must be worn when handling agents in the tissue culture room and the biosafety cabinet: –Latex or nitrile gloves –Lab coat –Safety glasses

18. Risk Assessment/Biosafety Level Engineering Controls [Modify list as appropriate] The following engineering controls must be used whenever handling agents in the tissue culture room: The following engineering controls must be used whenever handling agents in the tissue culture room: –Biosafety cabinet –Sink for Hand Washing –Mechanical pipettes –Sealed vials –Sharps containers/Broken Glass box –Eyewash/deluge shower (use only in an emergency)

19. Waste Disposal/Decontamination [Modify list as appropriate] Liquid waste will be treated with 10% household bleach and allowed to set for 20 minutes and drain-disposed. Solid red bag wastes will be placed in a red-bag lined bin and transferred to the _________ Building, Room______ medical waste storage area for pick up and disposal. Liquid waste will be treated with 10% household bleach and allowed to set for 20 minutes and drain-disposed. Solid red bag wastes will be placed in a red-bag lined bin and transferred to the _________ Building, Room______ medical waste storage area for pick up and disposal. Work areas must be disinfected with dilute bleach and 70% ethanol. Work areas must be disinfected with dilute bleach and 70% ethanol.

20. Emergency Procedures [modify as appropriate] In the event of a laboratory accident: In the event of a laboratory accident: –Tend to injured person –Notify the PI or the lab supervisor; inform the PI IMMEDIATELY if recombinant DNA was involved so that CLEB and the Biosafety Officer can be notified within 24 hrs. –Initiate clean up procedures. –Notify EHS, if the PI is not available, 642- 3073 or 642-3333 (After Hours) –Seek medical attention at Tang or the Alta Bates ER at 2450 Ashby Avenue after hours.

21. Conclusion This training is designed to inform you of the NIH requirements regarding use of recombinant DNA, and our lab’s implementation of the NIH requirements. All recombinant DNA experiments require BUA approval by CLEB and any changes require an approved amendment to the BUA. If you have any questions on this training, let me know, and I will contact the Biosafety Officer for assistance.