iGEM 2020 — Team Stanford

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# Team:Stanford/Notebook

Notebook

[![](https://static.igem.org/mediawiki/2020/3/37/T--Stanford--stanford-igem-logo.jpg)](https://2020.igem.org/Team%3AStanford)

<* Project

  [Motivation](https://2020.igem.org/Team%3AStanford/Description)
  [Description](https://2020.igem.org/Team%3AStanford/Description)
  [Parts](https://2020.igem.org/Team%3AStanford/Parts)
  [Contributions](https://2020.igem.org/Team%3AStanford/Contribution)
* Lab & Results

  [Safety](https://2020.igem.org/Team%3AStanford/Notebook)
  [Notebook](https://2020.igem.org/Team%3AStanford/Notebook)
  [Protocols](https://2020.igem.org/Team%3AStanford/Protocols)
  [Engineering Success](https://2020.igem.org/Team%3AStanford/Engineering)
* Computation

  [Model](https://2020.igem.org/Team%3AStanford/Model)
  [Software](https://2020.igem.org/Team%3AStanford/Software)
* Human Practices

  [Interviews](https://2020.igem.org/Team%3AStanford/Human_Practices)
  [Experiments](https://2020.igem.org/Team%3AStanford/Human_Practices)
  [Ethics](https://2020.igem.org/Team%3AStanford/Ethics)
* Future Steps

  [Implementation](https://2020.igem.org/Team%3AStanford/Implementation)
  [Applications](https://2020.igem.org/Team%3AStanford/Applications) [Entrepreneurship](https://2020.igem.org/Team%3AStanford/Entrepreneurship)
* Education

  [Speaker Series](https://2020.igem.org/Team%3AStanford/Education)
  [Lab Webinars](https://2020.igem.org/Team%3AStanford/Education)
  [Mentorship Program](https://2020.igem.org/Team%3AStanford/Education)
* People

  [Our Team](https://2020.igem.org/Team%3AStanford/Attributions)
  [Acknowledgements](https://2020.igem.org/Team%3AStanford/Attributions)
  [Attributions](https://2020.igem.org/Team%3AStanford/Attributions)
  [Collaborations](https://2020.igem.org/Team%3AStanford/Collaborations)
  [Citations](https://2020.igem.org/Team%3AStanford/Citations)

# Lab: Biosafety & Notebook

Our lab situation was a complicated one from the start. Due to the pandemic, we were not (and have never been) permitted onto Stanford's campus to use the Teaching Labs as we would have normally. While we could have scaled back our project to something either solely or heavily computational instead of choosing a project that requires substantial wet lab work to run, we were very motivated and excited for this project in particular, and decided we would do whatever it took to get into a lab space of some kind. Our quest to find lab space took us all over - from looking into outsourcing work to graduate students, to using a friend's garage lab, to automating the work through cloud labs such as [ECL](https://www.emeraldcloudlab.com/). Eventually, we ended up finding lab space in a biohackerspace / community lab in Santa Clara called [Biocurious](https://biocurious.org/), where four of our team members were able to work for some length of time.

## Biosafety at Biocurious

Prior to beginning work in BioCurious, all team members that worked at BioCurious had completed prior training for lab work on Stanford campus. All of our team members that worked in BioCurious had had existing certified training in General Safety, Biosafety, Life Science Safety, Chemical Safety, Compressed Gas Safety, and Bloodborne Pathogens Safety. Additionally, in order to begin work in BioCurious, the specific project our team was working on had to be approved by administrators at the lab. All work in BioCurious must be BSL-2 or lower to be approved to work in the space. Prior to beginning work, all members were required to have watched all safety training videos for BioCurious, as well as score a 100% on a required safety quiz prior to entering the building. For COVID-19 specific precautions, everyone working in BioCurious is required to wear a face mask at all times, remain 6 feet apart from other members, wipe down surfaces after use, and not reuse any disposables, including gloves. Members working at BioCurious are required to sign in, sign a safety acknowledgement, and no more than 15 people are allowed in the building at once. Non-members are not permitted entry to prevent the spread of COVID-19. While in lab, all of our researchers practiced all necessary safety precautions for dealing with bacteria, chemicals, nucleic acids, and compounds.

![Biocurious](https://static.igem.org/mediawiki/2020/9/97/T--Stanford--biocurious-banner.png)![](https://static.igem.org/mediawiki/2020/0/0f/T--Stanford--biocurious-safety.jpg)![](https://static.igem.org/mediawiki/2020/d/dd/T--Stanford--biocurious-gang.jpeg)

The organism we are working with (B. subtilis) is a generally-regarded-as-safe (GRAS), BSL-1, gram positive bacteria. Naturally found everywhere from the soil to the human gut microbiome, it is a non-pathogenic, extremely safe bacteria. When working with B. subtilis, our team still took all precautions, handling only with gloves, and adding bleach to cultures before disposing of them in a biohazardous waste container.  Any materials that may have come into contact with the bacteria were cleaned with bleach before disposal into a hazardous waste container. All genetic constructs ordered and used by the team contained no viral or pathogenic sequences. All chemicals used were disposed of properly in accordance with their safety sheet. Researchers wore gloves, long pants, and close-toed shoes at all times in lab. When necessary, researchers used lab coats, safety goggles, the fume hood, and alternate types of gloves.

## Considering Biosafety in Our Project

While developing our diagnostic, safety has been one of the most important considerations throughout its design, and was part of the reason that chose to use *Bacillus subtilis*. An ideal use case for our product involves at-home testing that can be performed by anyone, or even eventual integration into the human body as a biological threat monitor. When designing SEED, the health and safety of the user was at the forefront of our minds. We chose Bacillus subtilis due to its status as a GRAS, BSL-1 bacteria. It is already found naturally in the human microbiome, and is non-pathogenic. In an at home setting, accidental exposure to this bacteria should have very little risk to the user. For our homologous recombination detection method, it takes advantage of a natural system found in B. subtilis, meaning the risk of harmful side effects to the bacteria are low. Furthermore, by using a negative selection marker as the readout, cells that do not uptake the target are killed, and do not pose a threat to the user. Cells that do encounter the target and survive could have additional built in genetic mechanisms that will kill them and their lineage after a certain amount of time or when desired to prevent dangerous mutations. For the safety of our researchers, when performing all our experiments, B. subtilis was always kept on plates with antibiotics to prevent infection.

## Lab Notebook: Weekly Summaries

We have organized our lab notebook here by week instead of day. This is mostly because our lab work was sporadic and lagged behind at times, due to the way we had to receive our supplies. Instead of getting reagents and cells directly shipped to Biocurious for us to receive, everything had to be sent to our mentor's lab at Stanford and we had to coordinate with grad students to organize pickup times to come get what we had ordered. This bureaucratic process ultimately slowed the pace of our work.

#### Week 1: September 13-19

* Revived B. subtilis strains received from the Bacillus Stock Center (1A976 and 1A1276): Protocol #1
* Mini prepped pBS1C: Protocol #2
* Tested competence of 1A976: Protocol #4
* Took inventory and figured out what reagents we still needed to order
* Verified antibiotic resistances: We plated untransformed B. subtilis on kanamycin, chloramphenicol, and erythromycin plates as a negative control which. Plates were made by adding a 1000x dilution of the antibiotic stocks to standard LB plates.

![#](https://static.igem.org/mediawiki/2020/5/57/T--Stanford--Tubes.jpeg)

![#](https://static.igem.org/mediawiki/2020/d/db/T--Stanford--competent-b-sub.jpeg)

#### Week 2: September 20-26

* Attempted gibson assembly onto faulty pBS1C backbone: Protocol #5
* Transformed gibson results into E. Coli (got nothing): Protocol #6
* Tested competence of 1A976: Protocol #4

#### Week 3: September 27-October 3

* Continued trying to miniprep (pBS1C): Protocol #2
* Ran gels to confirm presence of pBS1C: Protocol #7
* Transformed miniprepped pBS1C into 1A976: Protocol #4
* Ran experiment to see if E. Coli were growing: we inoculated a 5mL tube containing nutrient broth with a colony from our E. Coli plate. We then allowed them to grow overnight and checked the OD in the morning

![#](https://static.igem.org/mediawiki/2020/2/2b/T--Stanford--gel-run.jpeg)

![#](https://static.igem.org/mediawiki/2020/a/a3/T--Stanford--MorePlates.jpeg)

#### Week 4: October 4-10

* Ordered new minipreps from Qiagen and Zymogen
* Ran experiment to test for sensitivity of DNA uptake by 1A976: we used Protocol #4, but adding differing amount of DNA to the cells (1 microgram, 500ng, 250ng, and 50ng of DNA)
* Got new cells containing pBS1C from the Bacillus Genetic Stock Center and mCherry and YFP plasmids from Dr. Elizabeth Libby of the Silver lab

#### Week 5: October 11-17

* Mini Prepped everything (mCherry plasmid, YFP plasmid, pBS1C): Protocol #2
* Gibson Assembly of ManP and Toehold Plasmids: Protocol #5
* Ran gel to test for assembly: Protocol #7
* Transformed manP and Toehold Plasmids into competent  E. Coli: Protocol #6

![GIBSON](https://static.igem.org/mediawiki/2020/0/04/T--Stanford--gibson.jpeg)

![#](https://static.igem.org/mediawiki/2020/1/1d/T--Stanford--AboveChris.jpeg)

#### Week 6: October 18-24

* Miniprepped E. Coli clones containing the assembled ManP and Toehold Plasmids in preparation for sequencing: Protocol #2. Tested ManP and Toehold constructsSent for sequencing via Elim BiopharmaceuticalsSelective plating on the antibiotic that they should have received resistance for via the assembled plasmids
* Miniprepped many colonies of pBS1C, mCherry plasmid, and YFP plasmid in preparation for large competence experiment: Protocol #2
* Tested competence by varying several parameters including time allowed for competence induction (1hr, 2hrs, and 3hrs), the temperature the cells grew at (37 degrees Celsius, and ~23 degrees celsius/room temperature), and agitation (cells were either shaken or not shaken): all combinations of these parameters were transformed via a protocol based on Protocol #4, but with requisite changes based on the variable conditions they were put under (i.e. if there were kept at room temperature, they were not put back into the shaker, but rather left on the bench at room temperature)
* Tested ManP and Toehold constructs;
  - Sent them for sequencing via Elim Biopharmaceuticals
  - Selective plating on the antibiotic that they should have received resistance for via the assembled plasmids

#### Week 7: October 25-Wiki Freeze

* Ran sporulation procedure: Protocol #8. Verified competence experiment from week 6 using the same procedure
* Verified competence experiment from week 6 using the same procedure

![#](https://static.igem.org/mediawiki/2020/4/4f/T--Stanford--chris-children.jpeg)

![Border](https://static.igem.org/mediawiki/2020/6/65/T--Stanford--border1.png)

##### about us

We are an interdisciplinary student team of eight undergraduates at Stanford University, mentored by Professors Stanley Qi and Drew Endy. Read more about [our team,](https://2020.igem.org/Team%3AStanford/Attributions)and contact us to learn more.
‍
**© Stanford iGEM 2020**

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![Stanford Medicine Logo](https://static.igem.org/mediawiki/2020/3/37/T--Stanford--stanford-medicine-logo.png)
![Biocurious Logo](https://static.igem.org/mediawiki/2020/8/82/T--Stanford--biocurious-logo.png)

![IDT logo](https://static.igem.org/mediawiki/2020/9/98/T--Stanford--idt-logo.png)
![BIOME logo](https://static.igem.org/mediawiki/2020/6/65/T--Stanford--biome-logo.png)

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# Team:Stanford/Engineering

Engineering

[![](https://static.igem.org/mediawiki/2020/3/37/T--Stanford--stanford-igem-logo.jpg)](https://2020.igem.org/Team%3AStanford)

<* Project

  [Motivation](https://2020.igem.org/Team%3AStanford/Description)
  [Description](https://2020.igem.org/Team%3AStanford/Description)
  [Parts](https://2020.igem.org/Team%3AStanford/Parts)
  [Contributions](https://2020.igem.org/Team%3AStanford/Contribution)
* Lab & Results

  [Safety](https://2020.igem.org/Team%3AStanford/Notebook)
  [Notebook](https://2020.igem.org/Team%3AStanford/Notebook)
  [Protocols](https://2020.igem.org/Team%3AStanford/Protocols)
  [Engineering Success](https://2020.igem.org/Team%3AStanford/Engineering)
* Computation

  [Model](https://2020.igem.org/Team%3AStanford/Model)
  [Software](https://2020.igem.org/Team%3AStanford/Software)
* Human Practices

  [Interviews](https://2020.igem.org/Team%3AStanford/Human_Practices)
  [Experiments](https://2020.igem.org/Team%3AStanford/Human_Practices)
  [Ethics](https://2020.igem.org/Team%3AStanford/Ethics)
* Future Steps

  [Implementation](https://2020.igem.org/Team%3AStanford/Implementation)
  [Applications](https://2020.igem.org/Team%3AStanford/Applications) [Entrepreneurship](https://2020.igem.org/Team%3AStanford/Entrepreneurship)
* Education

  [Speaker Series](https://2020.igem.org/Team%3AStanford/Education)
  [Lab Webinars](https://2020.igem.org/Team%3AStanford/Education)
  [Mentorship Program](https://2020.igem.org/Team%3AStanford/Education)
* People

  [Our Team](https://2020.igem.org/Team%3AStanford/Attributions)
  [Acknowledgements](https://2020.igem.org/Team%3AStanford/Attributions)
  [Attributions](https://2020.igem.org/Team%3AStanford/Attributions)
  [Collaborations](https://2020.igem.org/Team%3AStanford/Collaborations)
  [Citations](https://2020.igem.org/Team%3AStanford/Citations)

# Engineering

## What Does Engineering Success Look Like?

Science and engineering are processes that consist of constant learning and innovation. Successes in engineering are often incremental, as our team found this summer. Yet the small pieces of knowledge that you gather as you cycle through the engineering process accumulates later into larger successes. Thus, the important part of engineering is the process of designing, building, testing, and learning from the results.

#### Design

The first step of the cycle is designing possible solutions to a problem being presented. After this step, the engineer should have a blueprint laid for what they will build and test moving forward.

#### Build

In the next step, there should be some kind of physical product built and ready to be tested. In synthetic biology, this is often DNA constructs that are built and later tested.

#### Test

The third step is testing what has been designed and built; such as doing experiments in the lab. There should also be data gathered so that it can be analyzed and learned from afterwards.

#### Learn

The final step is to analyze the results from the testing phase, and find some takeaways from those results that can be learned from. This new knowledge is used to inform a new design step.

![DBT Cycle](https://static.igem.org/mediawiki/2020/5/5b/T--Stanford--dbt-cycle.png)

## Our Engineering Process

Our team spent a good chunk of our time throughout the iGEM season going through the engineering process from remote settings. We were not able to show a proof-of-concept for our technology because we were only able to get into lab during the end of the season, although we have begun that process. But the lack of lab space did not stop us from having successes throughout our journey.
‍

![Toehold Chart](https://static.igem.org/mediawiki/2020/f/f7/T--Stanford--engineering-toeholds.png)

The engineering of our toehold detection system began by parsing through existing literature to find and learn about RNA toeholds that have been verified to work in *E. coli* and cell free. After discovering several potential toeholds that had been tested in *E. coli*, our team modeled expression levels based on the RBS found in the loop of the toeholds. These toeholds and RBSs, optimized for *E. coli*, were not optimal for expression in *B. subtilis*. Not only do Bacillus subtilis use different RBSs, but the distance between the RBS and start codon must be different for *B. subtilis,* with around 7bp between them. With these differences in mind, we re-engineered our toeholds with optimal RBSs and spacing for *B. subtilis.*

Our original design plan was to target the coding sequence of cjBlue (BBa\_K592011), a blue chromoprotein. To create the toehold, we began by using the software developed by the [2017 Chinese University of Hong Kong iGEM team](https://2017.igem.org/Team%3AHong_Kong-CUHK/Software). Using their software, a user can input a target sequence, and it will return optimal target and toehold sequences, with additional estimates for binding affinities. After imputing the sequence for cjBlue, there did not appear to be any good toehold options with high binding affinities. So instead of using cjBlue as a target, our team switched to using the KanR gene. Targeting this gene would allow us to first verify expression of endogenous KanR by plating on kanamycin media to verify when the target is in the cell.

After settling on a toehold design and building the construct, we could then separate testing into two steps: testing the toehold in vivo with an endogenous target, then testing in conjunction with extracellular target uptake. To further verify the functionality of our toehold, we partnered with the 2020 Edinburgh iGEM team. We designed an experiment to be executed in cell free to test if the sequence could detect KanR. The toehold is coupled to sfGFP as a reporter, and targets a 36bp section of the KanR gene. The target will be introduced to the toehold in cell free, and if the toehold functions as intended, fluorescence can be measured with a plate reader.
‍

#### Next Steps

We plan to test more toeholds targeting various loci of the KanR gene, to determine what the ideal binding patterns are for B. subtilis. We will integrate this knowledge into our toehold software. Additionally, we will test expression levels of toeholds with alternate RBSs and spacing of the start codon to determine what is optimal for B. subtilis RNA toeholds. We will also determine the leakage of our toeholds in B. subtilis, and how to optimize their design and structure to minimize this noise.

![recbomination graph](https://static.igem.org/mediawiki/2020/d/d1/T--Stanford--recbomination-enginereing.png)

The engineering of our recombination detection system began by repurposing tools used for the miniBacillus project, developed by Wenzel and Altenbuchner (2015). Originally, the team ideated a system that would excise a transcription terminator, allowing transcription and translation of a reporter protein once recombination had occurred. Our team then created a model to see how much fluorescence would be visible, relative to the baseline with leakage. We discovered that when using a stronger promoter, fluorescence may be visible, but more leakage gives a higher baseline, making it harder to determine what is background and what is a signal. When using a weaker promoter, fluorescence would be too dim to allow for good functionality. Additionally, due to the rate of recombination, only a small number of cells would even begin to fluoresce in the first place, making the signal even harder to detect. Our solution to these problems was to change the type of readout we were building.

Instead of a colorimetric or fluorescent readout, we designed and engineered a system that would excise the negative selection marker ManP. In the presence of mannose, cells with the ManP cassette cannot survive. By flanking this gene with homology arms to our target sequence, the recombination triggered by the uptake of the target would remove the ManP gene, allowing cells that had encountered the target to survive in the presence of mannose. To test this new idea out, our team returned to modeling to determine the sensitivity of this detection method. We calculated that at least 1000 cells would be needed per test to reliably get colony growth in the presence of the target.

To further understand that rates of uptake and recombination, our team (now with access to lab space) designed and executed experiments to test the effectiveness of recombination into the B. subtilis genome. By using plasmids that integrate into the AmyE loci of the B. subtilis genome, any plasmids that successfully recombine into B. subtilis would fluoresce and gain kanamycin resistance, allowing for easy selection of transformed colonies. Our team was able to establish that inducibly-competent B. subtilis can be reliably transformed at 37°C with less than 100ng of DNA per 1x108 cells. Our next steps were then to vary the conditions at which we tested competence, such as incubation time, temperature, DNA concentration, and agitation. By varying these conditions, we were able to determine that recombination can occur after 1 hour of inducing competence and 1 hour of incubation with the plasmid DNA at 37°C. Recombination can occur with concentrations of DNA upwards of 300ng, and does not require agitation. Under ideal conditions, the rate of recombination is reliable enough for our system to function as intended.
‍

#### Next Steps

Now that we have proven that certain parts of the homologous recombination system would work, such as accurate signaling and natural competence in different conditions, we are working towards transforming the plasmid with the negative selection marker into our cells and then adding KanR to see if the system works.

![Competence Graph](https://static.igem.org/mediawiki/2020/0/03/T--Stanford--competence-engineering.png)

The competence of B. subtilis was an integral part of our diagnostic system design. It was essential that we verify the competence of our strains, and figure out how to optimize its induction for our desired use settings. To do this, we began by finding a basic protocol for transforming B. subtilis. This protocol had an incubation period to induce competence of 3 hours, and was performed at 37 degrees in an incubator shaker. After testing and retesting this protocol, we found that only the xylose-inducible strain 1A976 could be transformed in this manner.

To figure out how to induce competence for the mannitol-inducible strain 1A1276, we designed a new protocol for 1A1276. The cells would be incubated for 1 ½ hours before mannitol was added, then incubated for an additional 1 ½ hours after its addition. The mannitol was then removed before the DNA was added. Using this new protocol, we were able to successfully and reliably transform 1A1276.

After establishing a working protocol for both strains, our next step was to attempt to change variables in the protocol to optimize the test for an at home setting. To do this, we tried inducing competence without shaking, at room temperature and 37 degrees, without the addition of the inducing compounds, and for different incubation times. We found that competence could be induced without an incubator shaker after only an hour of incubation at 37 degrees. We were not able to get transformation at room temperature or without the addition of the inducing compound. However, we have been able to establish that competence could be induced using a household appliance like an oven, as shaking is not needed, and can be performed in a third of the traditional incubation time.

‍

#### Next Steps

We want our system to be able to be competent without the addition of sugar if possible, so we can allow for passive detection. We talk about the potential applications for that on our Applications Page. We are also planning to run experiments testing growth conditions and times at room temperature, also to look for potential passive uptake and detection.

## Our Engineering Successes

1. **Demonstrating High-Efficiency Transformation From Natural Competence**

Due to the COVID-19 pandemic, our team’s access to wet lab space was very limited. Because of this, we focused primarily on running experiments that would inform us on how effective a diagnostic system in B. Subtilis could be in an at-home setting. For us, the first step in this process was ensuring that we could reliably induce competence and transform B. subtilis using a simple procedure. We were able to do this in two cell lines, 1A976 and 1A1276, by simply adding sugar to their growth media to induce competence, then transforming them with ~1μg DNA. Both of these cell lines seemed to have very high transformation efficiency using this procedure as we were able to achieve lawn growth of the transformed cells after allowing them to grow overnight.

- **Testing Growth and Competence Conditions**

  The second step for us, was seeing what the limitations of the competence machinery in B. subtilis are. Specifically, we wanted to see what the limitations were surrounding transformation conditions (i.e. temperature, wait times, etc.) and amount of DNA needed for reliable transformation. To look for these limits, we ran a variety of experiments where we varied the temperature the cells were induced at (37C and room temperature), whether they were kept in a shaker, the amount of time given to induce competence (1hr, 2hr, and 3hr) and the amount of DNA added to the cells. From these experiments, we determined that competence can be achieved with less efficiency after only 1 hour of competence induction and ~400ng DNA added, and that reliable transformation can be achieved without the use of a shaker (but incubated at 37C) when ~400ng DNA was added.

![](https://static.igem.org/mediawiki/2020/0/05/T--Stanford--Poster_competence-table.png)

- **Validating Our Toehold Construct In Vitro**

  Throughout the competition, we had a very meaningful meaningful collaboration with the iGEM team at Edinburgh. In return for verifying their protocols, they offered to test out our toehold construct in vitro to verify that it works before we try transforming it into our cells to test in vivo. The result seen below shows that our toehold works and we designed it successfully.

![](https://static.igem.org/mediawiki/2020/d/d4/T--Stanford--Poster_toehold-proof.png)

![Curvy Border](https://static.igem.org/mediawiki/2020/6/65/T--Stanford--border1.png)

##### about us

We are an interdisciplinary student team of eight undergraduates at Stanford University, mentored by Professors Stanley Qi and Drew Endy. Read more about [our team,](https://2020.igem.org/Team%3AStanford/Attributions)and contact us to learn more.
‍
**© Stanford iGEM 2020**

![Bioengineering Logo](https://static.igem.org/mediawiki/2020/2/2e/T--Stanford--bioengineering-logo.png)
![Stanford Medicine Logo](https://static.igem.org/mediawiki/2020/3/37/T--Stanford--stanford-medicine-logo.png)
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# Team:Stanford/Experiments

# Experiments

Describe the research, experiments, and protocols you used in your iGEM project. These should be detailed enough for another team to repeat your experiments.

If you made Parts this year, please remember to put all information, characterization, and measurement data on the Part's Main Page on the [Registry](http://parts.igem.org/Main_Page).

### What should this page contain?

* Protocols
* Experiments

### Inspiration

* [2014 Colombia](https://2014.igem.org/Team%3AColombia/Protocols)
* [2014 Imperial](https://2014.igem.org/Team%3AImperial/Protocols)
* [2014 Caltech](https://2014.igem.org/Team%3ACaltech/Project/Experiments)

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:Stanford/Results

# Results

You can describe the results of your project and your future plans here.

### What should this page contain?

* Clearly and objectively describe the results of your work.
* Future plans for the project.
* Considerations for replicating the experiments.

### Describe what your results mean

* Interpretation of the results obtained during your project. Don't just show a plot/figure/graph/other, tell us what you think the data means. This is an important part of your project that the judges will look for.
* Show data, but remember **all measurement and characterization data must also be on the Part's Main Page on the [Registry](http://parts.igem.org/Main_Page).** Otherwise these data will not be in consideration for any medals or part awards!
* Consider including an analysis summary section to discuss what your results mean. Judges like to read what you think your data means, beyond all the data you have acquired during your project.

### Project Achievements

You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.

* A list of linked bullet points of the successful results during your project
* A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.

### Inspiration

See how other teams presented their results.

* [2014 TU Darmstadt](https://2014.igem.org/Team%3ATU_Darmstadt/Results/Pathway)
* [2014 Imperial](https://2014.igem.org/Team%3AImperial/Results)
* [2014 Paris Bettencourt](https://2014.igem.org/Team%3AParis_Bettencourt/Results)