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.