Team:Rutgers/Safety

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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Our experimental lab work this summer took place at the Waksman Institute of Microbiology on the UMDNJ campus at Rutgers University. This building is named in honor of Dr. Selman Waksman who was a soil microbiologist and a professor at Rutgers for four decades. In 1952, Dr. Waksman received the Nobel Prize for Physiology or Medicine for his discovery of streptomycin, which is isolated from <em>Streptomyces griseus</em>. This discovery led to his coining of the term &quot;antibiotic.&rdquo;<br />
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Following this discovery and the application of streptomycin as broad range antibiotic, questions regarding the resistance of bacteria in the presence of an antibiotic arose. Consequently, biological safety precautions had to be reevaluated to better reduce hazards and risks in the laboratory. &nbsp;<br />
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We believe that adherence to good laboratory practice is beneficial to many, not just the researcher. These qualities can help to avoid errors, allow for experimental procedures to be consistently repeated, and aids in the credibility of the work to others. Throughout the summer, we have carefully worked with <em>Escherichia coli</em> strain DH5a cells, various antibiotic media, chemical reagents and hazardous material. Our safety regulation and laboratory responsibilities included careful labeling schemes, detailed documentation and a cleaning routine of laboratory bench tops before and after work sessions. Waste collection and disposal for specific hazards was also carefully regulated.</span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Our iGEM projects involve the construction of gene regulatory networks that utilize positive and negative feedback loops found in microorganisms. The circuits we have proposed are made up of genetic devices such as regulators, repressors and reporters.<br />
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This is a template page. READ THESE INSTRUCTIONS.
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In the duration of our project we did not come across any DNA BioBricks that could cause pathogenic harm to the researchers, the public or the environment. &nbsp;Standard lab procedures require us to use Ethidium Bromide (EtBr), a florescent dye used to visualize DNA in agarose gels.</span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Rutgers Environmental Health and Safety regulates protocols and waste disposal methods used by federally funded research projects. &nbsp;(REHS, http://rehs.rutgers.edu/lsbio_comm.html)</span></span></p>
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You are provided with this team page template with which to start the iGEM season. You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki. You can find some examples <a href="https://2008.igem.org/Help:Template/Examples">HERE</a>.
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You <strong>MUST</strong> have a team description page, a project abstract, a complete project description, a lab notebook, and a safety page. PLEASE keep all of your pages within your teams namespace.
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Would any of your project ideas raise safety issues in terms of?</strong></span></span></h1>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">researcher safety</span></span></h1>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Use of hazardous chemicals: EtBr</span></span></h1>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">public safety</span></span>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">None unless mutated</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">environmental safety?</span></span>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">None unless mutated</span></span></li>
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<span class="Apple-style-span" style="font-family: arial, helvetica, sans-serif; font-size: 12px; "><strong>Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes,</strong></span><br />
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<span class="Apple-style-span" style="font-family: arial, helvetica, sans-serif; font-size: 12px; ">For the parts that are already in the registry no safety issues have been encountered. &nbsp;The two BioBrick parts we are creating from scratch are genetic regulators which should not raise any safety issues.</span></h1>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>3.&nbsp;Is there a local biosafety group, committee, or review board at your institution?</strong></span></span><span class="Apple-style-span" style="font-family: arial, helvetica, sans-serif; font-size: 12px; ">&nbsp;If yes, what does your local biosafety group think about your project?</span></h1>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">At Rutgers we have the department of Rutgers Environmental Health and safety they provide training for various hazards such as medical waste, hazardous waste, and radioactive waste. &nbsp;Dr Vershon our PI reviewed our project for possible safety issues, he did not find any. &nbsp;He also informed us of the labs procedures for disposing waste and general safety procedures.</span></span></h1>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>4.&nbsp;Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?</strong></span></span></h1>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">If extra money is available, Crystal Violet and other dyes are much safer and more accurate alternatives to EtBr.</span></span></h1>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Come up with an alternative to Bleach to clean out E.Coli &nbsp;from the sink and waterways.</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Use autoclavable/sterile materials instead of gratuitous amounts of plastic every day.<br />
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<strong>Software approach:</strong></span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>HDC - Hazardous DNA Compiler</strong></span></span>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Software that scans bricks for potentially hazardous DNA sequences</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Software that calculates risk of mutation that could create a potentially hazardous DNA sequence.</span></span></li>
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</ul>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Ethidium Bromide (EtBr) is used as a non-radioactive marker for identifying and visualizing nucleic acid bands in gel electrophoresis and in other methods of gel-based nucleic acid separation. EB is a dark red, crystalline, non-volatile solid, moderately soluble in water, which fluoresces readily with a reddish-brown color when exposed to ultraviolet light. Its formula is 2,7,-Diamino-10-ethyl-9-phenyl-phenanthridium bromide,</span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Hazards</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">EtBr is a <strong>potent mutagen</strong> and moderately toxic after an acute exposure. Since EtBr can be absorbed through skin and considered to be an irritant to the skin, eyes, mouth, and upper respiratory tract, it should be handled with gloves. &nbsp;EtBr should be stored away from strong oxidizing agents in a cool, dry place, and the container must be kept undamaged and tightly closed.</span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Safety Precautions</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Pure EB should only be handled in a fume hood, with the user wearing protective equipment that includes a lab coat, closed-toe shoes, chemically resistant gloves, and chemical safety goggles (not just safety glasses). &nbsp;Nitrile gloves are recommended for short term exposure to EB. &nbsp;After use, users should wash hands and discard gloves. &nbsp;When using UV light to activate the EB, be sure to use the appropriate</span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>General Laboratory Rules</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">No eating, drinking, smoking, applying cosmetics, etc. in the lab. &nbsp;</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Access to the laboratory is restricted to only when the Dr. Vershon, Dr. Mead, Dr. Gelfand or Ms. Chen from the Vershon lab is present. &nbsp;</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Please do not bring friends into the lab (Dr. Vershon has our roster. If we add new members we will okay it with him before they are allowed in the lab).</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Work areas should be cleaned after any chemical spill.</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Mechanical pipeting devices are always used. &nbsp;<em>Mouth pipeting is not permitted.</em></span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Hands should be washed before and after working in the laboratory.</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Disposable gloves should be worn when working with hazardous chemicals such as ethidium bromide. &nbsp;Lab coats may be worn in the lab to protect your street clothes from contamination. &nbsp;Lab coats should not be worn outside of the laboratory. &nbsp;</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Shorts and sandals are not permitted in the lab.</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Waste must be disposed of properly (see Waste Disposal on p.1-5).</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Sterility/aseptic techniques</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Pipette tip boxes and mf tube bags must always be <u>tightly</u> closed.</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Autoclaved bottles must always be capped <u>tightly</u>.</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Common courtesy</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">If you are using Dr. Vershon&rsquo;s supplies (i.e. media, plates, reagents) always inform someone when supplies are very low. <u>Don&rsquo;t take the last bottle</u>!</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Help out making media when requested</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Sterile solutions should not be returned</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><em>(Source: iGEM-Lab1-Pipeting; Vershon, 2011)</em></span></span></p>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Storage and saving</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Incubator (30 </strong><strong>o</strong><strong>C)</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Plates (agar side up, properly labeled and documented in the lab notebook!)</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Liquid cultures (tightly capped, properly labeled and documented in the lab notebook!)</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><u>Turn on shaker</u>after turning off!</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Turn off light</span></span></li>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Room temperature (20 </strong><strong>o</strong><strong>C - 25 </strong><strong>o</strong><strong>C)</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">DNA Kit (with lyophilized DNA)</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Ethanol</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Concentrated Buffers</span></span></li>
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</ul>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Fridge (4 </strong><strong>o</strong><strong>C)</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Plates (agar side up, properly labeled and documented in the lab notebook!)</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Liquid cultures (tightly capped, properly labeled and documented in the lab notebook!)</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">DNA (i.e. un-lyophilized DNA)</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Buffers</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Media</span></span></li>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Freezer (-20 </strong><strong>o</strong><strong>C)</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Most restriction enzymes (always read the label!)</span></span></li>
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</ul>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Low-temperature Freezer (-80 </strong><strong>o</strong><strong>C)</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Frozen cultures (properly labeled and documented in the lab notebook!)</span></span></li>
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</ul>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; <strong>Light sensitive</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Tetracycline is light sensitive and should be saved in the fridge with foil around it and tape.</span></span></li>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">If a bottle is brown, it is light sensitive and should stay this way.</span></span></li>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Cleaning up</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Generally, make sure everything in the lab is in order. Cover things so they may not collect dust. Always close pipet tip boxes and bags with mf tubes to maintain their sterility. Keep autoclaved bottles <u>tightly</u> closed.</span></span></p>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Glassware. </strong>All tubes must be washed out in Waksman Room 234. Place the glass tube and the cap in its appropriate basket.</span></span></p>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Disposables</strong>. If it&rsquo;s made out of plastic and you do not need it, it&rsquo;s probably garbage. Make sure before throwing out tubes that you are absolutely sure you don&rsquo;t need it. If you are continuing someone elses experiment and/or you are not sure whether to save the tube, save, label and freeze it just in case.</span></span></p>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Fridge/freezers.</strong>All fridge/freezer contents must be labeled properly and documented in the lab notebook. Tape plates together if they are related. Try not to tape only one side of the stack because if it falls apart or just falls, it gets messy. Reuse tape when you can and again, label the tape. All tubes must be capped tightly.</span></span></p>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>Biohazard (<em>E. coli)</em></strong>. Add (a small amount) of chlorine bleach to waste containers and let it sit for a bit before discarding it down the sink.</span></span></p>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">The drawers are now labeled with respect to their contents, please keep them this way. The window should be shut completely if you have opened it. Place all books back on the shelf. Place all lab notebooks and protocols in the brown binder. Wipe the bench with some ethanol at the end of the day.</span></span></p>
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&nbsp;</p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;"><strong>References</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Barker, Kathy. <em>At The Bench: A Laboratory Navigator</em>. New York: Cold Spring Harbor Laboratory, 1998. Print.</span></span></p>
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<p style="margin-left:.5in;">
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Rastogi, Smita, and Neelam Pathak. <em>Genetic Engineering</em>. New Delhi: Oxford UP, 2009. Print.</span></span></p>
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<p style="margin-left:.5in;">
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Seidman, Lisa A., and Cynthia J. Moore. <em>Basic Laboratory Methods for Biotechnology: Textbook and Laboratory Reference</em>. San Francisco: Pearson/Benjamin Cummings, 2009. Print.</span></span></p>
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Nelson, David L., Albert L. Lehninger, and Michael M. Cox. <em>Lehninger Principles of Biochemistry</em>. New York: W.H. Freeman, 2008. Print.</span></span></p>
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<p style="margin-left:.5in;">
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<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Vershon, Andrew, Barth Grant, Bryce Nickels, and Janet Mead. <em>Introduction to Research in Genetics Laboratory Manual, Spring 2011</em>. Print.</span></span></p></body>
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!align="center"|[[Team:Rutgers|Home]]
 
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!align="center"|[[Team:Rutgers/Team|Team]]
 
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!align="center"|[https://igem.org/Team.cgi?year=2010&team_name=Rutgers Official Team Profile]
 
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!align="center"|[[Team:Rutgers/Project|Project]]
 
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!align="center"|[[Team:Rutgers/Parts|Parts Submitted to the Registry]]
 
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!align="center"|[[Team:Rutgers/Modeling|Modeling]]
 
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!align="center"|[[Team:Rutgers/Notebook|Notebook]]
 
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!align="center"|[[Team:Rutgers/Safety|Safety]]
 
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!align="center"|[[Team:Rutgers/Attributions|Attributions]]
 
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==Safety==
 
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Please use this page to answer the safety questions posed on the [[Safety | safety page]].
 

Revision as of 01:15, 15 July 2011

Our experimental lab work this summer took place at the Waksman Institute of Microbiology on the UMDNJ campus at Rutgers University. This building is named in honor of Dr. Selman Waksman who was a soil microbiologist and a professor at Rutgers for four decades. In 1952, Dr. Waksman received the Nobel Prize for Physiology or Medicine for his discovery of streptomycin, which is isolated from Streptomyces griseus. This discovery led to his coining of the term "antibiotic.”
Following this discovery and the application of streptomycin as broad range antibiotic, questions regarding the resistance of bacteria in the presence of an antibiotic arose. Consequently, biological safety precautions had to be reevaluated to better reduce hazards and risks in the laboratory.  
We believe that adherence to good laboratory practice is beneficial to many, not just the researcher. These qualities can help to avoid errors, allow for experimental procedures to be consistently repeated, and aids in the credibility of the work to others. Throughout the summer, we have carefully worked with Escherichia coli strain DH5a cells, various antibiotic media, chemical reagents and hazardous material. Our safety regulation and laboratory responsibilities included careful labeling schemes, detailed documentation and a cleaning routine of laboratory bench tops before and after work sessions. Waste collection and disposal for specific hazards was also carefully regulated.</p>

Our iGEM projects involve the construction of gene regulatory networks that utilize positive and negative feedback loops found in microorganisms. The circuits we have proposed are made up of genetic devices such as regulators, repressors and reporters.
In the duration of our project we did not come across any DNA BioBricks that could cause pathogenic harm to the researchers, the public or the environment.  Standard lab procedures require us to use Ethidium Bromide (EtBr), a florescent dye used to visualize DNA in agarose gels.

Rutgers Environmental Health and Safety regulates protocols and waste disposal methods used by federally funded research projects.  (REHS, http://rehs.rutgers.edu/lsbio_comm.html)

  1. Would any of your project ideas raise safety issues in terms of?

    • researcher safety

      • Use of hazardous chemicals: EtBr

    • public safety
      • None unless mutated
    • environmental safety?
      • None unless mutated


Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes,

  • For the parts that are already in the registry no safety issues have been encountered.  The two BioBrick parts we are creating from scratch are genetic regulators which should not raise any safety issues.


3. Is there a local biosafety group, committee, or review board at your institution? If yes, what does your local biosafety group think about your project?

  • At Rutgers we have the department of Rutgers Environmental Health and safety they provide training for various hazards such as medical waste, hazardous waste, and radioactive waste.  Dr Vershon our PI reviewed our project for possible safety issues, he did not find any.  He also informed us of the labs procedures for disposing waste and general safety procedures.


4. Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

  • If extra money is available, Crystal Violet and other dyes are much safer and more accurate alternatives to EtBr.

  • Come up with an alternative to Bleach to clean out E.Coli  from the sink and waterways.
  • Use autoclavable/sterile materials instead of gratuitous amounts of plastic every day.
    Software approach:
  • HDC - Hazardous DNA Compiler
    • Software that scans bricks for potentially hazardous DNA sequences
    • Software that calculates risk of mutation that could create a potentially hazardous DNA sequence.



Ethidium Bromide (EtBr) is used as a non-radioactive marker for identifying and visualizing nucleic acid bands in gel electrophoresis and in other methods of gel-based nucleic acid separation. EB is a dark red, crystalline, non-volatile solid, moderately soluble in water, which fluoresces readily with a reddish-brown color when exposed to ultraviolet light. Its formula is 2,7,-Diamino-10-ethyl-9-phenyl-phenanthridium bromide,

Hazards

EtBr is a potent mutagen and moderately toxic after an acute exposure. Since EtBr can be absorbed through skin and considered to be an irritant to the skin, eyes, mouth, and upper respiratory tract, it should be handled with gloves.  EtBr should be stored away from strong oxidizing agents in a cool, dry place, and the container must be kept undamaged and tightly closed.

Safety Precautions

Pure EB should only be handled in a fume hood, with the user wearing protective equipment that includes a lab coat, closed-toe shoes, chemically resistant gloves, and chemical safety goggles (not just safety glasses).  Nitrile gloves are recommended for short term exposure to EB.  After use, users should wash hands and discard gloves.  When using UV light to activate the EB, be sure to use the appropriate

General Laboratory Rules

  1. No eating, drinking, smoking, applying cosmetics, etc. in the lab.  
  2. Access to the laboratory is restricted to only when the Dr. Vershon, Dr. Mead, Dr. Gelfand or Ms. Chen from the Vershon lab is present.  
  3. Please do not bring friends into the lab (Dr. Vershon has our roster. If we add new members we will okay it with him before they are allowed in the lab).
  4. Work areas should be cleaned after any chemical spill.
  5. Mechanical pipeting devices are always used.  Mouth pipeting is not permitted.
  6. Hands should be washed before and after working in the laboratory.
  7. Disposable gloves should be worn when working with hazardous chemicals such as ethidium bromide.  Lab coats may be worn in the lab to protect your street clothes from contamination.  Lab coats should not be worn outside of the laboratory.  
  8. Shorts and sandals are not permitted in the lab.
  9. Waste must be disposed of properly (see Waste Disposal on p.1-5).
  10. Sterility/aseptic techniques
  11. Pipette tip boxes and mf tube bags must always be tightly closed.
  12. Autoclaved bottles must always be capped tightly.
  13. Common courtesy
  14. If you are using Dr. Vershon’s supplies (i.e. media, plates, reagents) always inform someone when supplies are very low. Don’t take the last bottle!
  15. Help out making media when requested
  16. Sterile solutions should not be returned

(Source: iGEM-Lab1-Pipeting; Vershon, 2011)

 

Storage and saving

Incubator (30 oC)

  • Plates (agar side up, properly labeled and documented in the lab notebook!)
  • Liquid cultures (tightly capped, properly labeled and documented in the lab notebook!)
  • Turn on shakerafter turning off!
  • Turn off light

 

Room temperature (20 oC - 25 oC)

  • DNA Kit (with lyophilized DNA)
  • Ethanol
  • Concentrated Buffers

 

Fridge (4 oC)

  • Plates (agar side up, properly labeled and documented in the lab notebook!)
  • Liquid cultures (tightly capped, properly labeled and documented in the lab notebook!)
  • DNA (i.e. un-lyophilized DNA)
  • Buffers
  • Media

 

Freezer (-20 oC)

  • Most restriction enzymes (always read the label!)

 

Low-temperature Freezer (-80 oC)

  • Frozen cultures (properly labeled and documented in the lab notebook!)

           

            Light sensitive

  • Tetracycline is light sensitive and should be saved in the fridge with foil around it and tape.
  • If a bottle is brown, it is light sensitive and should stay this way.

 

Cleaning up

Generally, make sure everything in the lab is in order. Cover things so they may not collect dust. Always close pipet tip boxes and bags with mf tubes to maintain their sterility. Keep autoclaved bottles tightly closed.

 

Glassware. All tubes must be washed out in Waksman Room 234. Place the glass tube and the cap in its appropriate basket.

 

Disposables. If it’s made out of plastic and you do not need it, it’s probably garbage. Make sure before throwing out tubes that you are absolutely sure you don’t need it. If you are continuing someone elses experiment and/or you are not sure whether to save the tube, save, label and freeze it just in case.

 

Fridge/freezers.All fridge/freezer contents must be labeled properly and documented in the lab notebook. Tape plates together if they are related. Try not to tape only one side of the stack because if it falls apart or just falls, it gets messy. Reuse tape when you can and again, label the tape. All tubes must be capped tightly.

 

Biohazard (E. coli). Add (a small amount) of chlorine bleach to waste containers and let it sit for a bit before discarding it down the sink.

 

The drawers are now labeled with respect to their contents, please keep them this way. The window should be shut completely if you have opened it. Place all books back on the shelf. Place all lab notebooks and protocols in the brown binder. Wipe the bench with some ethanol at the end of the day.

 

References

Barker, Kathy. At The Bench: A Laboratory Navigator. New York: Cold Spring Harbor Laboratory, 1998. Print.

Rastogi, Smita, and Neelam Pathak. Genetic Engineering. New Delhi: Oxford UP, 2009. Print.

Seidman, Lisa A., and Cynthia J. Moore. Basic Laboratory Methods for Biotechnology: Textbook and Laboratory Reference. San Francisco: Pearson/Benjamin Cummings, 2009. Print.

Nelson, David L., Albert L. Lehninger, and Michael M. Cox. Lehninger Principles of Biochemistry. New York: W.H. Freeman, 2008. Print.

Vershon, Andrew, Barth Grant, Bryce Nickels, and Janet Mead. Introduction to Research in Genetics Laboratory Manual, Spring 2011. Print.

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