iGEM 2016 — Team Dundee_Schools

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# Team:Dundee Schools/Notebook

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# Dundee Schools

#### Notebook

## Wet Team

#### Week 1 - Familiarising ourselves with the labs

* Transformation + Overnight of pSBIC3
* Mini Prep of pSBIC3
* Restriction Digest of pSB1C3

##### PCR’s:

+ Hfq *E. coli*- Unsuccessful
+ Hfq *Serratia*- Unsuccessful
+ OsmY- Unsuccessful

* Mini Prep of pSBIC3
* Restriction Digest of pSB1C3

#### Week 2 - Carrying on with cloning

##### PCR’s:

+ Hfq *E. coli*- Unsuccessful
+ Hfq *Serratia* – Unsuccessful
+ osmY- Unsuccessful

##### 3rd PCR set up:

+ Hfq *E. coli* – Successful
+ Hfq *Serratia* – Unsuccessful
+ OsmY - Successful

##### PCR’s:

+ Hfq *Serratia* 2 at 49°C - Successful
+ Hfq *Serratia* 1 at 65°C - Successful

* Ligation into PSBIC3 and transformed
* The 3 successful PCR’s were plated at concentrations of 2:1 and 3:1 with vector; no plates grew colonies

#### Week 3 - Made our first BioBrick

* Redid all 3 PCR’S – Hfq *E. coli* and osmY both worked. Hfq *Serratia* did not.
* Colony PCR of the successful PCR’s, overnight, mini prep, sent away for sequencing and then stocked.
**First 2 BioBricks made: OsmY + Hfq *E. coli**** PCR of *osmY-hfq E. coli* fusion and PCR of *osmY-hfq Serratia* Fusion - successful

+ Digested and Ligated etc.… sequenced then stocked.

**2 BioBricks made: osmY Hfq *E. coli* Fusion + osmY Hfq *Serratia* Fusion**

#### Week 4 - Started to make gene fragments

* PCR of Hfq *Serratia* - successful

+ Sequenced and Stocked.

**1 Bio brick made: Hfq *Serratia**** Gene fragments – EC-SRNA and sma-SRNA digested and ligated into pSBIC3
* Digestion of Rhamnose promoter plasmid

#### Week 5 – Started to add Ha tags

##### Ligations:

+ rbs-osmY-Hfq-*E. coli*
+ rbs-osmY-Hfq-*Serratia*
+ rbs-osmY-Hfq-*E. coli*- Ha Tag
+ rbs-osmY-Hfq-*Serratia*- Ha Tag

##### PCR’s:

+ rbs-osmY- Ha Tag - Successful
+ rbs-osmY-Hfq *E. coli*- Ha Tag- Successful
+ rbs-osmY-Hfq *Serratia*-Ha Tag – Successful

* All PCR’s transformed and overnighted.

#### Week 6 - Phage outbreak

##### Overnights of:

+ rbs-osmY-Hfq- Ha tag
+ rbs-osmY-Hfq *E. coli*- Ha tag
+ rbs-osmY-Hfq *Serratia*- Ha tag

* Mini preps, sequencing and transformations of the above
* Phage outbreak on Thursday; due to this all transformations were lost, but re-done the next day
* All sent away for sequencing
* Ec – SRNA and sma SRNA – both stocked.

#### Week 7 - Western blots

* We did our first western blot test with no timed samples.
* We also completed a timed western blot by adding different concentrations of Rhamnose and taking samples every hour

##### Primers:

+ *fliC* for Ec-sRNA
+ *virF* for Ec-sRNA

#### Week 8 - Final week

* Performed another Western Blot to get a much cleaner image for the whole cell
* Overnights with Ec-SRNA *fliC* in MG1655 and Ec-SRNA *virF*
* Plates grew colonies – colony PCR with colonies.
* Sent away for sequencing

## Dry Team

#### Week 1 -

* Started off discussing our topic of bacterial infections and brainstorming how we could target them; settled on RNA interference.
* Split the team into 2 groups, wet team (Matthew, Bartosz, Albert) and dry team (Mia, Beth, Darryl).
* Decided what each team had to do for that week
* Started with general research on our chosen infections (Cholera and Shigellosis)
* Darryl started and completed learning basic HTML and CSS via tutorials on the site Codecadamy, then began the initial stages of the wiki so as to get to grips with it.

#### Week 2 -

Began to contact various people/organisations:

+ Colalife
+ Mercy Ships
+ Water Aid
+ Lifeline Express

* Had our first meeting about maths modelling and discussed how we could incorporate it into our project.
* Further developed wiki.

#### Week 3 -

* Had a phone call with Brian Walshe of Mercy Ships

Started organisation for Northern meet up:

- Began developing presentation content and layout
- Work began on poster

#### Week 4 -

* Spoke to Rob Porter (Prof. of clinical microbiology) via email.
* Planned a skype call with Simon Berry of Colalife.
* Contacted British Red Cross.

#### Week 5 -

* Attended the Edinburgh Northern meet up & presented our project there, to which we were met with excellent feedback.
* Carried on working in the lab and progressing with our presentation.
* Further development of poster.

#### Week 6 -

* Contacted British Pharmaceutical Society
* Contacted Mark McCulloch (Army Doctor)
* Carried on with making our poster for the UK meet up.

#### Week 7 -

* Set up meeting with Mark
* Finished poster
* Carried on with our wiki content
* Received our FBI sunglasses and pens for the Jamboree.

#### Week 8 -

* Attended the UK meet up in London – presented our talk and our poster to multiple UK iGEM teams.
* Got our hoodies and t-shirts for Boston.
* Had our agent photoshoot.

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