Always remember that we are working with hazardous materials. Sloppy handling of such materials does not only endanger you, but also other course participants.
The Howard Hughes Medical Institute provides safety data (click on Select an LCSS) for the most commonly used chemicals in modern research labs. Quick and easy to use. VWR provides a more extensive list of MSDSs (Material Safety Data Sheets) - it may be worth having a look at those if you are in doubt. There are also links to MSDSs from other manufacturers or institutions here. For quick reference, consult Appendix 12 in Sambrook & Russel, vol 3 (available in the lab).
1. No eating or drinking in the laboratory. Do not store food or drink in any of the laboratory refrigerators.
2. All accidents and spillages must be reported to the instructor or the TA.
3. Lab coats, goggles and gloves have to be worn while working with hazardous materials such as acids or the chemicals below. Open-toe sandals should not be worn while handling these materials. The general use of lab coats and closed shoes is strongly recommended.
4. Extreme caution must be taken while handling or in the presence of the following hazardous agents in the lab:
a. Ethidium bromide (EtBr). This is known mutagen and presumed carcinogen that can be absorbed through the skin. Always wear gloves and check for leaks before handling EtBr. Handle EtBr only in designated areas, and stain gels only in designated containers. Dispose of EtBr contaminated materials in designated containers. Any spills of EtBr-containing solutions must be wiped up immediately, and spill areas monitored with a hand-held UV illuminator following clean-up. See Molecular Probes MSDS.
b. SybrGreen - although SybrGreen is much safer than EtBr, it still binds to DNA (otherwise it wouldn't detect DNA), and thus should be treated with caution. You should therefore use the same protection as for EtBr. SybrGreen also comes in a solution containing DMSO (Molecular Probes MSDS), which is toxic.
c. Acrylamide is one of the nastiest chemicals we have. It has moderate acute toxicity, but high chronic neurotoxicity. Be extremely careful when using it - use only in fumehood, wear lab coat, goggles and gloves. Acrylamide is less dangerous if polymerized - but watch out for unpolymerized lquid. Spillages may dry out and produce dust which can be breathed in - if you spill anything, inform the instructors. Never leave a spillage. See also MSDS of BioRad, the manufacturer of our acrylamide.
d. TEMED, a reagent used to polymerize acrylamide, is very nasty and easily produces toxic fumes (see Stratgene MSDS). Only use in fumehood.
e. Ultraviolet (UV) transilluminators (room MAR 169). Protective goggles or face shield must be worn before turning on any UV light source. Exposure must be kept to a minimum. Avoid leaning over the transilluminator or leaving the transilluminator on longer than necessary to capture the image on the gel documentation system.
f. Phenol and chloroform. Phenol/chloroform extractions must be done in the fume hood only, and wastes must be disposed of in designated containers only. Both nitrile and latex gloves provide only some protection from phenol (see glove manufacturers recommendations), so be careful not to spill on your gloves, and if you do, change gloves immediately. See also MSDS for phenol and chloroform.
g. Live cultures. Contaminated wastes must be disposed of into biohazard waste containers only. Nondisposable wares used for cultures must be placed in designated trays for autoclaving.
h. Sharps (needles, broken glass, Pasteur pipettes, etc). Disposal must be into designated containers only. EtBr-contaminated sharps must be segregated. Do not recap needles!
5. Make sure all gas burners and hot plates are turned off after use. Do not spray alcohol to disinfect any area while the gas burner is on!
6. All nondisposable nonbiohazard glasswares must be rinsed before leaving in soaking trays in the sink.
7. Take gloves off from hands before touching any fixtures in the lab (e.g. telephones, door knobs, computer keyboards, common equipment unless designated otherwise, books etc.) whether or not you think the gloves are clean.
8. Do not pipette solutions by mouth.
9. Wash hands thoroughly before leaving the laboratory.
1. Do not move any equipment, or change any set parameters on any equipment, without permission from instructor or TA.
2. Follow operating instructions where posted on or near equipment - that means you have to read them! If you have any doubt at all about proper operation of any piece of equipment, consult instructor or TA.
3. Report any equipment problems or malfunctions right away. Don't just use an alternative method or equipment.
4. Pipetman micropipettes. Do not exceed the maximum and minimum settings on the pipettes. Failure to observe this rule could result in damage to the pipettes and/or alter their calibration.
| Type |
Minimum setting
|
Maximum setting
|
| P1000 |
100 ul
|
1000 ul
|
| P200 |
10 ul
|
200 ul
|
| P100 |
5 ul
|
100 ul
|
| P20 |
1 ul
|
20 ul
|
| P10 |
0.5 ul
|
10 ul
|
Never lay down a pipette on the bench with the tip on - any liquid in the tip may flow into the pipette barrel and cause damage.
5. Gel electrophoresis equipment and power supplies. Observe maximum allowable setting for each power supply. Always turn the voltage/current adjustment control to minimum before turning the power supply on or off. Always turn power off before opening or touching the electrophoresis chamber.
6. UV transilluminator. Must be turned on only with UV shield/goggles on. The top must be cleaned with 95% ethanol and distilled water after each use.
1. Store all samples and solutions that need refrigeration in the refrigerator/freezer space designated to you. Materials that are stable at room temperature can be stored at your lab work station.
2. All samples and solutions must be properly labeled. In all cases proper labeling includes name of owner and identity/nature of sample/solution. Solutions should also be labeled with the date on which they are prepared. Improperly labeled materials may be discarded.
3. All enzymes must be kept on ice or in refrigerator boxes on the lab bench. Use only a fresh sterile pipette tip for every aliquot taken from the enzyme vial. Enzymes must be handled gently and promptly returned to the freezer or proper storage after use. Restriction enzymes should be taken out of the freezer only immediately before use.
4. Frozen enzyme stock stolutions (e.g. Proteinase K) must be thawed on ice.