iGEM 2019 — Team Unimelb

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# Team:Unimelb/Notebook

Unimelb iGEM

* [Home](https://2019.igem.org/Team%3AUnimelb)
* [Team](https://2019.igem.org/Team%3AUnimelb/TeamSummary)
* [Project](https://2019.igem.org/Team%3AUnimelb/ProjectSummary)

* [Human Practices](https://2019.igem.org/Team%3AUnimelb/Human_PracticesSummary)
* [Awards](https://2019.igem.org/Team%3AUnimelb/AwardsSummary)
* [Judging Form](https://igem.org/2019_Judging_Form?team=Unimelb)

![Unimelb Team Logo](https://static.igem.org/mediawiki/2019/0/01/T--Unimelb--logoBanner.png)

## Notebook

**JUNE:**

04/06/2019

* Created a list of deliverables and experimental plans for next month - Sam
* Made sure all the new members have completed their laboratory induction training - Antonio
* Outreach team discussed and finalised events for the month of August and September – Lydia, Emily, Betty
* Followed up on the student grant which we had previously applied - Emily

05/06/2019

* Meeting with our advisor regarding our project plan and outreach events - Everyone

**JULY:**

05/07/2019

* Created protocols for cell-competency test, transfection and ligation – Sam
* Discussed potential problems and solutions regarding the final product of our project – Lab team
* Discussed on how we want to take our project in the future – Lab team

09/07/2019

* Ethics poll created and shared via social media by outreach team – Lydia

16/07/2019

* Wiki collaboration with US AFRL CarrollHS Lab Pats – Xavier

17/07/2019

* Had a skype chat with University of Sydney iGEM team regarding both of our projects and discussed about potential collaboration – Sam, Xavier

25/07/2019 & 26/07/2019

* Attended a two-day winter workshop organised by our advisor. We gained useful insights on Project management, Outreach and Social media strategy for iGEM, how to integrate ELSI into our iGEM project and Fundraising strategy as well – Everyone

30/07/2019

* Created an elaborate lab experimental workflow to get feedback from our mentor - Sam

**AUGUST:**

1/08/2019

* Preparation of KCl and MgCl2 – Aish, Sam, Thanushi

7/08/2019

* Preparation of SOC medium and LB agar –

8/08/2019

* Preparation of Agar plates with Ampicillin and Chloramphenicol – Sam, Aish

18/08/2019

* Outreach team had a stall during Open day and explained to the public regarding iGEM and our project.
* They had a poll with 4 different scenarios and understood the public opinions on synthetic biology and how we could incorporate it in our project write up – Lydia, Betty, Emily, Sam

20/08/2019

* Meeting with our Advisor regarding our progress and open day ethics review – Everyone
* Drafted emails for our potential sponsors – Antonio, Xavier
* Discussion about Synthetic Biology Australasia (SBA) conference and who is going to represent the team - Everyone

28/08/2019

* Met with our mentor to finalise and clarify any doubts regarding our experimental plan – Sam, Antonio

**SEPTEMBER:**

5/09/2019

* Preparation of LB medium – Aish, Sam
* Attended Bioethics conference for a period of three days which gave us a way to engage with the local ethics community - Lydia

10/09/2019

* Created rough template for our website and write up for the WIKI – Xavier
* Ordered the IDT sequences and reagents – Sam
* Attended a talk on “Stem cells – From bench to Bedside” – Aish, Thanushi

13/09/2019

* Cell lysis and measurement of iGluSnFr activity – Antonio, Thanushi
* The cell did not lyse by ultra-sonification. So, we had to look at other ways to lyse them.

19/09/2019

* Cell competency test – Aish, Antonio

20/09/2019

* Counting colonies and calculating efficiency from the previous cell competent test – Xinran

25/09/2019

* The outreach group organised a two-day event for young kids to inspire them to learn science through hands-on and fun filled activities – Lydia, Betty, Emily, Aish, Xinran, Xavier, Thanushi

**OCTOBER:**

1/10/2019

* Preparing clonal colonies for IDT genes which included digestion of gene fragments, ligation of gene fragments into plasmid and transformation – Sam, Antonio, Thanushi, Aish
* Assigning people to fill out the judging criteria – Sam, Aish, Antonio, Thanushi

3/10/2019

* The transfections did not work. We did not find any cells. Troubleshooting involved loading and running a gel electrophoresis – Sam, Thanushi, Aish, Antonio
* The gel did not work as expected making us to repeat the entire cloning process.

4/10/2019

* Checking point of failure for cloning


[![](https://2019.igem.org/wiki/images/8/8b/HQ_page_logo.jpg)](https://2019.igem.org)

# Team:Unimelb/Experiments

Unimelb iGEM

* [Home](https://2019.igem.org/Team%3AUnimelb)
* [Team](https://2019.igem.org/Team%3AUnimelb/TeamSummary)
* [Project](https://2019.igem.org/Team%3AUnimelb/ProjectSummary)

* [Human Practices](https://2019.igem.org/Team%3AUnimelb/Human_PracticesSummary)
* [Awards](https://2019.igem.org/Team%3AUnimelb/AwardsSummary)
* [Judging Form](https://igem.org/2019_Judging_Form?team=Unimelb)

![Unimelb Team Logo](https://static.igem.org/mediawiki/2019/0/01/T--Unimelb--logoBanner.png)

## Experiments

**SOC MEDIUM**

* To 950 mL of deionised water, add:
  Tryptone 20g
  Yeast Extract 5g
  NaCl 0.5g
* Dissolve the solutes by placing the flask on the shaker
* Add 10 mL of a 250 mM solution of KCl
* Add 5 mL of sterile solution of 2 M MgCl2 once solution is 60°C or less
* Fill to 980 mL with deionised water
* Autoclave the SOC medium at 121°C for 20 minutes for sterilisation
* Add 20 mL of sterile 1 M solution of glucose
* Sterilise by passing it through a 0.22-µm filter
* Store the SOC medium at room temperature and for long term storage, store at 4°C.

**LB MEDIUM (1L LIQUID)**

* For 1L LB medium, the correct amounts are:
  NaCl 10g
  Peptone (or tryptone) 10g
  Yeast extract 5g
* Collect them in in a bottle and add 1L of dH2O
* Autoclave the LB medium at 121°C for 20 minutes for sterilisation.

**LB AGAR**

* For 200 mL LB-agar, the correct amounts are:
  NaCl 2g
  Peptone (or tryptone) 2g
  Yeast extract 1g
  Bacto-agar 3g
* Collect them in a bottle and add 200 mL of H2O
* Autoclave the LB medium at 121°C for 20 minutes for sterilisation.

Note: With 200 mL LB-agar 8 plates can be made.

**AMPICILLIN STOCK (100 mg/ml)**

* Dissolve 0.5 g ampicillin in 5 mL Milli Q.
* Mix/vortex so that all the ampicillin goes into solution.
* Filter into a falcon tube using a syringe and a 0.22 μm filter for sterilization.
* Aliquot into smaller Eppendorf tubes.
* Store at -20°C under dark conditions.

 **CHLORAMPHENICOL STOCK (25mg/ml)**

* Dissolve 0.125 g chloramphenicol in 5 mL Milli Q.
* Mix/vortex so that all the ampicillin goes into solution.
* Filter into a falcon tube using a syringe and a 0.22 μm filter for sterilization.
* Aliquot into smaller Eppendorf tubes.
* Store at -20°C under dark conditions.

**CELL COMPETENCY TEST**

For cell competency test we used iGEM Competent Cell Test Kit according to the instructions provided by [http://parts.igem.org/Help:2018\_Competent\_Cell\_Test\_Kit](http://parts.igem.org/Help%3A2018_Competent_Cell_Test_Kit) .

[![](https://2019.igem.org/wiki/images/8/8b/HQ_page_logo.jpg)](https://2019.igem.org)

# Team:Unimelb/Results

Unimelb iGEM

* [Home](https://2019.igem.org/Team%3AUnimelb)
* [Team](https://2019.igem.org/Team%3AUnimelb/TeamSummary)
* [Project](https://2019.igem.org/Team%3AUnimelb/ProjectSummary)

* [Human Practices](https://2019.igem.org/Team%3AUnimelb/Human_PracticesSummary)
* [Awards](https://2019.igem.org/Team%3AUnimelb/AwardsSummary)
* [Judging Form](https://igem.org/2019_Judging_Form?team=Unimelb)

![Unimelb Team Logo](https://static.igem.org/mediawiki/2019/0/01/T--Unimelb--logoBanner.png)

## Results

Here you can describe the results of your project and your future plans.

### 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.** 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)