Week 15 Pre-Regionals

9/28/13-10/3/13

With the wiki frozen and the North American Regional Jamboree looming, we focused on getting all of our data together for the presentation and poster. Refinement of our presentation and poster were our top priorities before heading off to Toronto.

10/4/13-10/6/13

Time for the North American Regional Jamboree! We were glad to have our presentation early on Saturday, so we could relax and enjoy the other presentations. Everyone did an incredible job, and Toronto was a blast. We are thrilled to have advanced to the World Championship Jamboree and cannot wait to go to MIT.

Week 16 Post-Regionals

10/7/13-10/12/13

After taking a brief break to catch up on some sleep and schoolwork, we met with our advisors to discuss what we plan to accomplish before the World Championships. We decided on expanding our RiboTALs to targeting GFP under the control of the well characterized Anderson Promoter Collection. Since the Anderson Promoters are a family of constitutive promoters that have measured relative strengths, we believe they can give us predictable system responses. We plan to use 5 of the Anderson Promoters: J23100, J23101, J23105, J23106 and J23109, which have a wide range of activity and use our model to predict the response of the other Anderson Promoters. We will also continue to improve and populate the Depot.

Week 17

10/13/13-10/19/13

We're back to doing more construction as we try to replace the pTet promoter in front of TBS2+GFP with our chosen Anderson Promoters. After a few issues, we managed to transform the promoters from the iGEM distribution kits and use Standard Assembly to assemble them together with TBS2+GFP which we PCR amplified out of our previous constructs. Our Standard Assemblies were successful, so we did contransformations of the Anderson Promoters+TBS2+GFP in pSB1A2 and our Riboswitch2+TAL8 RiboTAL in pSB3K3. We had hoped to start testing our new constructs in the Tecan on Thursday, but due to the lack of/slow growing colonies for our cotransformations testing will have to be held off. Tecan testing will start next week, due to other unforeseen circumstances.

Week 18

10/20/13-10/26/13

This week was full of Tecan runs. With all of our constructs contransformed and stored as glycerol stocks, it was just of matter of getting them all onto plates and finding time to run them in the Tecan for 8-10 hours. All of our runs were done in triplicate with 6 different arabinose concentrations and 5 different theophylline concentrations. Our run schedule went something like this:

Monday	J23100+TBS2GFP// R2T8
Tuesday	J23109+TBS2GFP// R2T8 and J23105+TBS2GFP// R2T8
Wednesday	J23106+TBS2GFP// R2T8 and J23101+TBS2GFP// R2T8
Thursday	J23106+TBS2GFP// R2T8
Friday	J23105+TBS2GFP// R2T8

Not all of our runs gave us satisfactory data, so some were redone. Overall our data is looking good. A lot of work is now going into analyzing our data and using it to improve our model.