Team:Penn/Safety

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<title>University of Pennsylvania iGEM</title>
<title>University of Pennsylvania iGEM</title>
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<a href="https://2014.igem.org/Team:Penn"><li>Home</li></a>
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  <li>Project
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  <ul>
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      <a href="https://2014.igem.org/Team:Penn/Overview"> <li>Overview</li> </a>
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    <a href="https://2014.igem.org/Team:Penn/Magnetism"> <li>Magnetism</li> </a>
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    <a href="https://2014.igem.org/Team:Penn/Microbio"> <li>Microbiology</li> </a>
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    <a href="https://2014.igem.org/Team:Penn/Synbio"> <li>SynBio in AMB-1</li> </a>
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    <a href="https://2014.igem.org/Team:Penn/CdTolerance"> <li>Cadmium Tolerance</li> </a>
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  <li>Human Practices
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      <a href="https://2014.igem.org/Team:Penn/Specsheet"><li>Spec Sheet</li></a>
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    <a href="https://2014.igem.org/Team:Penn/Outreach"><li>Outreach</li></a>
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    <a href="https://2014.igem.org/Team:Penn/Biomeme"><li>Biomeme</li></a>
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</li>
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  <li>Notebook
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  <ul>
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      <a href="https://2014.igem.org/Team:Penn/Notebook"><li>Timeline</li></a>
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      <a href="https://2014.igem.org/Team:Penn/Safety"><li>Safety</li></a>
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    <a href="https://2014.igem.org/Team:Penn/Protocol"><li>Protocols</li></a>
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    <a href="https://2014.igem.org/Team:Penn/Supplement"><li>Supplementary Materials</li></a>
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    <a href="https://2014.igem.org/Team:Penn/Resources"><li>Resources</li></a>
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  </ul>
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  </li>
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  <li>Team
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      <a href="https://2014.igem.org/Team:Penn/Team"><li>About Us</li></a>
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    <a href="https://2014.igem.org/Team:Penn/Sponsors"><li>Sponsors</li></a>
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<div style = "text-align: center; font-size: 24px;">Timeline</div></br>
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<h3>Biological chassis</h3>
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<p>Our work throughout iGEM has been split between using three different E. coli strains (DH5 alpha, W3110, and MG 1655) and Magnetospirillum Magneticum AMB-1. All four of these strains pose minimal risk inside the lab. We use strict biosafety laboratory techniques such as wearing gloves when handling these organisms and any piece of lab ware that comes in contact with them.
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<h3>E. coli</h3>
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<p> All three strains of E. coli are highly conserved and pose minor risks when handled carefully in the lab. They are all rod shaped, gram negative bacteria. While E. coli survives in the intestinal tract of many organisms, all three strains used in the lab are safe to work with as they lack the adaptability and capability to cause infection. ATCC has assigned all strains a biosafety level of 1. In order to prevent contamination of any samples in the lab, including mammalian cell cultures, we dispose of all E. coli by bleaching cultures and then disposing of equipment in biohazard. We also wear gloves and exercise caution while handling anything with bacteria.
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</p>
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<h3>Magnetospirillum Magneticum AMB-1</h3>
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<h5><u><b> AMB-1 in the Lab </b></u></h6>
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<h3 style= "text-align: center;">Biological chassis</h3>
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<p> Magnetospirillum Magneticum AMB-1 is a strain of magnetic bacteria that poses minimal health risk to humans in a lab environment. This strain of bacteria is too sensitive to environmental conditions to cause infections to humans. At the same time, all procedures used to handle E. coli were also used with Magnetospirillum Magneticum AMB-1. This includes bleaching old cultures, disposing old tubes according to biohazard protocols and wearing gloves while handling cultures.
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<ol>
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</p>
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  <li>Molecolar Biology Training Workshop</li>
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    <li>Practiced the basics of molecular cloning</li>
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</ol>
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<h3 style= "text-align: center;">Week 2</h3>
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    <li>Idea Brainstorming and Generation</li>
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    <li>Compiled a preliminary list of potential ideas</li>
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</ol>
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<h3 style= "text-align: center;">Week 3</h3>
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    <li>Settled on two main ideas: 1. Quorum sensing with antibiotics 2. Heavy metal removal with Magnetotactic bacteria</li>
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    <li>Learned how to use Geneious to design cloning process</li>
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    <li>Practiced extracting biobricks and transforming NEB Turbo cells with plasmids</li>
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    <li>Human Practices</li>
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<h5><u><b> AMB-1 in the Environment </b></u></h3>
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    <li>Visited Biomeme to get the portable qPCR machine</li>
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</ol>
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<h3 style= "text-align: center;">Week 4</h3>
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<ol>
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    <li>Decided on the project idea: Heavy metal removal with Magnetotactic bacteria
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    <li>Identified primers and promoters in AMB-1 strain
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    <li>Identified AMB-1 transformation vector
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    <li>Extracted smtA biobrick gene and made glycerol stock
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</ol>
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<h3 style= "text-align: center;">Week 5</h3>
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    <li>Developed three goals to accomplish for the project
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    <li>Determined the constructs to clone into AMB-1
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    <li>Designed two fast-fail experiments
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  <li> Created a workflow for the construction of plasmid
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  <li> Human Practices
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<p> The use of AMB-1 as a chassis for bioremediation also eliminates some safety concerns that synthetic biologists had with E.Coli. Using an engineered strain of E.Coli runs the risk of disrupting natural ecosystems through competition of species and horizontal gene transfer. This major safety dilemma is addressed by creating a foundational chassis that can be removed from the contaminated water by manipulating its magnetic properties. Implementation of a filtration device incorporating this bioremediation agent in a water treatment facility (add link to robot page) would allow us to remove cadmium from the water with minimal impact on the aquatic environments. This is also a serious safety improvement from chemical precipitation and ion exchange methods that are currently used to treat cadmium pollution because it does not leave harmful byproducts that could negatively impact the organisms that populate the waterways.</p>
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    <li>Planned outreach events at high school summer programs
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<h3 style= "text-align: center;">Week 6</h3>
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    <li>Ordered chemicals for AMB-1 growth medium
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    <li>Completed primer design
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    <li>Obtained AMB-1 strain from Dr.Goolian
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<h3 style= "text-align: center;">Week 7</h3>
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<ol>
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    <li>Determined the OD600 plate reading conversion formola experimentally
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    <li>Attempted to do E.Coli and AMB-1 cadmium tolerance test
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    <li>Designed all construct on Geneious
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    <li>Human Practices
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<h3> Public Safety </h3>
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    <li>Met with Dr.Rizk to discuss presenting to incoming bioengineering freshmen
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<h3 style= "text-align: center;">Week 8</h3>
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    <li>Ordered DNAs from Genscript and IDT
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<h3 style= "text-align: center;">Week 9</h3>
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    <li>Redid E.Coli Cadmium Tolerance Test in M9 media
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    <li>Learned to do cell count for AMB-1
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    <li>Attempted to make AMB-1 chemically competent
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    <li>Human Practices
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<p> Since some of our bacteria may have antibiotic resistance and could theoretically pose a health risk, we use biohazard protocols to prevent any release of our bacteria from the lab. This includes bleaching cultures before disposing of them and disposing of any old tubes in biohazard containers. This eliminates the risk of any bacteria escaping our lab or making their way into any adjacent labs. If released accidentally, none of the strains we used in our project generate any toxic proteins and should not have harmful impacts. </p>
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    <li>Presented to high school students at Penn M&T program
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    <li>Planned a synthetic biology preceptorial
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    <li>Contacted Schuylkill Action Network
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<h3 style= "text-align: center;">Week 10</h3>
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    <li>Completed AMB-1 growth curve under different media
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    <li>Determined cell count and OD600 conversion formola for AMB-1 experimentally
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    <li>Completed E.Coli Cadmium Tolerance Test in LB media
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    <li>Human Practices
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<h3> Safety Training  </h3>
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  <li> Volunteered at SEA Science Carnival
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<h3 style= "text-align: center;">Week 11</h3>
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    <li>Addressed EMSGM growth problem with pH
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    <li>Make new recovery media with adjusted pH for optimal AMB-1 growth
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    <li>Chemical Transformation with different recovery broths
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    <li>Attempted PCR assembly with synthesized parts
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<h3 style= "text-align: center;">Week 12</h3>
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    <li>Used anaerobic chambers to grow plates
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    <li>Troubleshooted lack of magnetism with new iron maleate solution
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    <li>Troubleshooted AMB-1 electroporation experiment with plating at every step
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    <li>Began cloning successfolly assembled PCRed constructs into PYMB essentials
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<h3 style= "text-align: center;">Week 13</h3>
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    <li>Finished cloning flurescent protein with smtA and mCherry into PYMB essentials
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    <li>Transform AMB-1 with this construct to test for successfol transformation with fluorescence test
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<h3 style= "text-align: center;">Week 14</h3>
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    <li>Make new E-MSGM media
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    <li>Re-designed construct
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<h3 style= "text-align: center;">Week 15</h3>
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    <li>Troubleshooted cloning E. coli construct for cadmium tolerance
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    <li>Sequence verified finished constructs on PYMB
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<h3 style= "text-align: center;">Week 16</h3>
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<ol>
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    <li>Built the prototype of a portable spectrophotometer for cell recovery experiment
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    <li>Designed primers for Gibson Assembly of final construct
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    <li>Human Practices
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<p> All team members have participated in a thorough Penn environmental and safety training course that helped us prepare for work with hazardous substances, infectious agents and human source materials. A link to this course is provided below. Also, a link to the national biosafety guidelines has also been provided.  
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    <li>Presented to Penn BE 100 lecture to intriduce synthetic biology to freshman bioengineers.
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Penn lab safety course: http://www.ehrs.upenn.edu/programs/bio/  
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</ol>
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National biosafety guidelines: http://www.absa.org/resbslinks.html
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<h3 style= "text-align: center;">Week 17</h3>
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</p>
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<ol>
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    <li>Tested AMB-1 aerobic colture and anaerobic colture with magnetometer
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    <li>Troubleshooted and retried cloning E. coli construct
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    <li>PCRed up parts we received for construction of shuttle vector
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</ol>
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<h3 style= "text-align: center;">Week 18</h3>
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    <li>Tested AMB-1 aerobic colture and anaerobic colture with magnetometer
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    <li>Finished PCRing parts we received from IDT
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</ol>
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<h3 style= "text-align: center;">Week 19</h3>
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<ol>
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  <li> Measured the magnetic strength indicator T2 for various concentration of AMB-1 colture and attempted to develop a trendline
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  <li> Counted folly grown sample of aerobic AMB-1 and made a trendline converting OD to cell concentration
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    <li>Cloned all biobricks into PSB1C3 backbone
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</ol>
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<h3 style= "text-align: center;">Week 20</h3>
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<ol>
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    <li>compiled all data and graphics for wiki
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</ol>
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</ol></div>
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Latest revision as of 01:15, 18 October 2014

University of Pennsylvania iGEM

Biological chassis

Our work throughout iGEM has been split between using three different E. coli strains (DH5 alpha, W3110, and MG 1655) and Magnetospirillum Magneticum AMB-1. All four of these strains pose minimal risk inside the lab. We use strict biosafety laboratory techniques such as wearing gloves when handling these organisms and any piece of lab ware that comes in contact with them.

E. coli

All three strains of E. coli are highly conserved and pose minor risks when handled carefully in the lab. They are all rod shaped, gram negative bacteria. While E. coli survives in the intestinal tract of many organisms, all three strains used in the lab are safe to work with as they lack the adaptability and capability to cause infection. ATCC has assigned all strains a biosafety level of 1. In order to prevent contamination of any samples in the lab, including mammalian cell cultures, we dispose of all E. coli by bleaching cultures and then disposing of equipment in biohazard. We also wear gloves and exercise caution while handling anything with bacteria.

Magnetospirillum Magneticum AMB-1

AMB-1 in the Lab

Magnetospirillum Magneticum AMB-1 is a strain of magnetic bacteria that poses minimal health risk to humans in a lab environment. This strain of bacteria is too sensitive to environmental conditions to cause infections to humans. At the same time, all procedures used to handle E. coli were also used with Magnetospirillum Magneticum AMB-1. This includes bleaching old cultures, disposing old tubes according to biohazard protocols and wearing gloves while handling cultures.

AMB-1 in the Environment

The use of AMB-1 as a chassis for bioremediation also eliminates some safety concerns that synthetic biologists had with E.Coli. Using an engineered strain of E.Coli runs the risk of disrupting natural ecosystems through competition of species and horizontal gene transfer. This major safety dilemma is addressed by creating a foundational chassis that can be removed from the contaminated water by manipulating its magnetic properties. Implementation of a filtration device incorporating this bioremediation agent in a water treatment facility (add link to robot page) would allow us to remove cadmium from the water with minimal impact on the aquatic environments. This is also a serious safety improvement from chemical precipitation and ion exchange methods that are currently used to treat cadmium pollution because it does not leave harmful byproducts that could negatively impact the organisms that populate the waterways.

Public Safety

Since some of our bacteria may have antibiotic resistance and could theoretically pose a health risk, we use biohazard protocols to prevent any release of our bacteria from the lab. This includes bleaching cultures before disposing of them and disposing of any old tubes in biohazard containers. This eliminates the risk of any bacteria escaping our lab or making their way into any adjacent labs. If released accidentally, none of the strains we used in our project generate any toxic proteins and should not have harmful impacts.

Safety Training

All team members have participated in a thorough Penn environmental and safety training course that helped us prepare for work with hazardous substances, infectious agents and human source materials. A link to this course is provided below. Also, a link to the national biosafety guidelines has also been provided. Penn lab safety course: http://www.ehrs.upenn.edu/programs/bio/ National biosafety guidelines: http://www.absa.org/resbslinks.html