LABORATORY CLASSES SCHOOL OF BIOLOGICAL SCIENCES NISER 1 INDEX PAGE 1. Laboratory Safety guidelines 3-10 2. Laboratory Protocols for odd semester A. First Semester: Biology Laboratory 11-33 B. Third Semester: Microbiology Laboratory 34-61 C. Third Semester: Biochemistry laboratory 62-79 D. Fifth Semester: Animal physiology laboratory 80-93 E. Fifth Semester: Plant Physiology Laboratory 94-133 F. Seventh Semester: Developmental Biology/Genetics: received on 30.4.2013 265-280 G. Seventh Semester: Genetic engineering: received on 30.4.2013 281-288 3. Laboratory protocols for even semester A. Second Semester: Biology Laboratory 134-157 B. Fourth Semester: Cell biology Laboratory 158-180 C. Fourth Semester: Genetics Laboratory 181-218 D. Sixth Semester: Immunology Laboratory 219-225 E. Sixth Semester: Molecular Biology Laboratory 226-264 F. Seventh semester: Bioinformatics: protocol pending 2 Laboratory Safety Guidelines School of Biological Sciences NISER, BBSR, India The School of Biological Sciences, NISER is committed to providing a safe environment for all. However, laboratory safety is a mutual responsibility and requires full participation and cooperation of all involved persons - students, researchers, and faculty and staff members. The following Lab Safety Guidelines have been established for your protection as a student in the school of biological sciences. These rules will be rigidly and impartially enforced. Noncompliance may result in a grading penalty and/or dismissal from lab. 3 Personal Protection 1. Safety glasses must be worn at all times in the Lab. This is the policy of NISER and a national safety requirement. The glasses must be of the impact protection type with splash guards and must meet required specifications. Other eye/face protection may be required with specific procedures. The laboratory in-charge has approved safety glasses. The glasses must be worn at all times within the lab space - even while working on the computer or writing in your lab book. 2. Contact lenses are discouraged. The safety of wearing contact lenses in laboratories has been hotly debated over the last several years. NISER is preparing guidelines indicating that contact lenses can be worn if and only if proper protective eyewear is also worn. The school of biological sciences recognizes that some eye conditions require contacts for certain vision correction therapies. However, students who choose to wear contacts must recognize the inherent increased risks - they are difficult to remove if chemicals get in the eye, they have a tendency to prevent natural eye fluids from removing contaminants, and sudden displacement can cause visual problems that create additional hazards. Soft contact lenses are especially problematic because they can discolor and also absorb chemical vapors causing damage before the wearer is alerted to the problem. If you choose to wear contacts, please tell your lab instructor and individual will be liable for any personal damages caused as a result of wearing the contact lens. 3. Appropriate gloves will be provided when needed. Use of gloves is required for handling certain chemicals and biological. Gloves are very expensive. Do not change gloves needlessly. For some uses double gloving may be necessary, your instructor will guide you for such need. 4. Appropriate clothing is required. Your clothing is a barrier between your skin and chemicals. You must be covered to the knee - also no bare midriffs or shoulders. Knee length shorts are acceptable, anything above the knee - shorts, skirts, or dresses are not. Lab coats are. 5. Shoes must be worn. No sandals, thongs, open toed or open heeled shoes. 6. Roll up sleeves and tie up loose clothing and long hair when working with equipment, open flame, any chemicals or biological substances. 7. Do not eat, drink (including sport bottles and water bottles), or store food in the labs. 8. Smoking or use of other tobacco products is prohibited. 9. Wash hands after working with chemicals. 10. It is the recommendation of this department that all students of reproductive age, especially women who have recently conceived or are anticipating conception 4 during the semester, discuss the course content and reagents with their physician if they are concerned about reproductive toxins. General Lab Rules 1. Conduct yourself in a responsible manner at all times in the laboratory. 2. When first entering a lab room, do not touch any equipment, chemicals, or other materials in the laboratory area until you are instructed to do so. 3. Read all instructions carefully and plan your work. Understand the experiment and if in doubt, ask. 4. Follow the written lab procedure - laboratory activity at this level is not meant to be creative. Improper combinations or amounts of chemicals can be very dangerous. No unauthorized experiments are to be performed. If you are curious about trying a procedure not covered in the experimental procedure, consult with your laboratory instructor. 5. Lab tables should be as uncluttered as possible to allow work space and avoid accidents. Also, keep the aisles clear to prevent tripping over your gear, and so that other people can pass unhampered. Place book bags, pocketbooks, etc. under the lab tables. In some labs, seats or stools are not to be used during labs - students need to be mobile to avoid possible spills and are not to place themselves under the edge of the lab bench where chemicals may spill. 6. Lab activities require your undivided attention. No music allowed in student labs. Radios (including Walkman type) and other entertainment devices are not permitted. No mobile phone use. 7. Lab computers are for laboratory business only - no unrelated Internet surfing or checking e-mails. 8. Treat chemicals with respect and understand the chemicals you are using. Material Safety Data Sheets (MSDSs) are available in the binders in each room or consult your instructor or the laboratory manager to find them. Do not remove the MSDSs from the binders. Bring the binder to the Biology Lab Manager to request a copy. 9. Learn where the safety and first-aid equipment is located. This includes fire extinguishers, fire blankets, and eyewash stations. 10. Notify the instructor immediately in case of an accident, no matter how small it seems. 11. Students are never permitted in the Biology storage rooms or preparation areas unless given specific permission by their instructor or head of the department/institution. 12. Handle all living organisms used in a laboratory activity in a humane manner. Preserved biological materials are to be treated with respect and disposed of properly. 5 13. Leave the lab area clean. Put equipment and chemicals away and wipe off the bench top. Laboratory Usage Education: At a glance • Know your laboratory well before you actually start working. • Know functioning/handling of all instruments before actually using them. • Know what you should not do with the instrument. • Enter into logbooks of instruments after each use. • Know the chemicals that you will be handling in each of your study. • Keep the glassware and plastic wares you need for your study at your disposal well ahead of your actual study time. • Know the nature and handling of chemicals/reagents that you will be handling. • Know the dos and do-not in the lab before functioning. • Know the trouble shooting of instruments well. • Know the handling of the waste you generate from your study. • Keep record of your day to day activities in a record book where you enter all information: positive/negative. • Inform your PI about your progress of study time to time. 6 LAB DOS AND DON’TS LAB DOS LAB DON’TS 1. Know the location of all safety and emergency equipment used in the lab. 2. Know use of fire extinguishers and their locations. 1. NEVER experiment on your own. 3. Know the location of the closest telephone. 2. Do not eat or drink in the lab room at any time. 4. Familiarize yourself with all lab procedures before 3. Do not chew gum or eat candy during lab doing the lab exercise. exercises. 5. Report ALL accidents, hazards or chemical spills 4. NEVER add water to an acid or vice-versa. to the instructor (no matter how small). 5. Do not wear contacts in the lab without proper eye 6. Keep you work area clean and clutter free. protection. 7. Tie back all long hair and remove dangling jewelry 6. NEVER smell, taste or touch chemicals. during lab. 7. NEVER work in the lab alone. 8. Always be sure that electrical equipment is turned in the "off" position before plugging it into a socket. 8. NEVER use electrical equipment around water. 9. Handle all animals with care 9. NEVER mix chemicals before asking the instructor. 10. Use extreme care when handling sharp objects. 10. NEVER return unused chemicals to the original container. 11. Dispose of all chemicals, broken glass and other lab materials into the proper containers as directed 11. Absolutely NO PLAY is allowed in the lab area! by the instructor. 12 NEVER leave the lab area without washing your 12. When heating liquids in a test tube, always point hands the test tube away from anyone. 13. Keep all materials away from open flames. 7 Disposal of Wastes: 1. Do not dispose off chemicals in the sink. (Rule of Thumb: If you don’t want to drink it, don’t dump it in the sink). Follow your instructor’s directions for disposal. Be sure to dispose of chemicals in the proper waste collector. Do not mix chemical waste without being instructed to do so. Any container that is used to collect chemical waste must be properly labeled and closed at all times unless actively pouring into it. 2. Properly dispose of animal tissue in the MARKED Biohazard bags. Never throw animal tissue in lab garbage cans.