Team:William and Mary/Notebook


NOTEBOOK
Week 1 (May 29-June 4)
Wetlab Summary
Starting off the summer by cloning parts that will enable us to test the basic functionality of the mf-Lon for protein degradation. These parts consist of pBad and J231** parts for all of the sfGFP pdt tag variants, as well as mf-Lon. Currently we have confirmed versions of 3 of 6 J231** sfGFP variants, as well as an mf-Lon under the control of J231**.
Outreach Summary
We began to lay down the foundation for our big outreach project, which will become a searchable database of past outreach projects from award-winning teams. Project categories are being brainstormed in addition to tags for different audiences.
Week 2 (June 5-June 11)
Wetlab Summary
Continued to make progress towards functionally validating and testing our mf-Lon and pdt parts. While our pBad sfGFP pdt tags have been lagging behind, we moved ahead with making combination parts for our simple circuit (LacI pdt and pLac sfGFP pdt combos), and we have been validating our existing mf-Lon and pdt tagged sfGFPs. From the current data, it appears that some of our sequence confirmed pdts are working, while others are not. However, all of our mf-Lons are functional.
Outreach Summary
We entered information about 2016 and 2015 IHP/EPE award-winners and decided to move the database information to a spread sheet to make it easier to eventually create a website. Alyssa met with Professor Griffin, who specializes in community-based research and outreach. We worked on the PowerPoint and brochure for our Bioengineering Lab, and the PowerPoint is now playing and visible to anyone passing by our lab. GSCC got back in touch with Alyssa on possibly organizing a field trip for Girl Scouts to the BEL. In addition to that, Alyssa communicated with Kelley Clark from the STEM Education Alliance to organize education activities with AP Bio/9th Grade Bio students in the fall.
Week 3 (June 12-June 18)
Wetlab Summary
This week, we were held up a great deal due to not being able to send for sequencing. We are currently in experiments to confirm pdts are working, and cloning inducible sfGFP pdt parts to be able to measure speed. We are switching over to pLac instead of pBad, which entails additional concerns due to the need for a repressor.
Outreach Summary
Alyssa got in touch with Jacqueline from GSCC to discuss doing activities with the Girl Scouts. We will host about 50 girls on two Saturdays either in September or October. For the database, we've gone back to 2011 outreach projects but will likely end up going back to 2008. Alyssa started organizing a speaker series, and three professors have agreed to speak. Theresa and Alyssa started working on a video for our project. Sejal finished updating the K-5 and 6-8 activity booklets.
Week 4 (June 19-June 25)
Wetlab Summary
We finally confirmed an mf-Lon and pdt combo. We currently have characterization of J23100 tags along with a pBad mf-Lon. We are also trying to functionally confirm a J23116 mf-Lon, and characterize R0011 (pLac0-1) pdts with pBad mf-Lon. One issue we have been running into is with mf-lon causing toxicity/growth inhibition. We will aim to tune our parts around this.
Outreach Summary
We set up the dates for the GSCC visit in October. We have continued to enter entries into the database spreadsheet and have decided to house the database in the wiki so it will be accessible to other iGEM teams and iGEM leadership. We made and edited the script for the viedo describing our project. Alyssa recorded the voiceover and Theresa filmed the footage.
Week 5 (June 26-July 2)
Wetlab Summary
This week we focused mainly on two things. Number one is continuing to validate and clone parts. Number two is determining exactly how we plan to measure everything. The first one has made steady progress, (cloning of combination parts, quorum circuits, and functional validation of our pLac pBad circuit), while the second one still requires a great deal more thinking. Namely we have to investigate some claims from a paper, as well as figure out more robustly what we want to measure.
Outreach Summary
We published the video explaining our project to YouTube and Facebook. We now have two dates locked down for our bioengineering speaker series. Professor Schniepp is speaking on the 13th and Professor Kerscher is speaking on the 20th (4:00-5:00). Sejal and Alyssa continued working on the high school synthetic biology activities booklet.
Week 6 (July 3-July 9)
Wetlab Summary
This week we were focused on trying to measure a functional induction curve with a "steady state" as well as confirming our pdts at the single cell level. We managed to accomplish the latter, though the effect size is not perfect, and there is still a lot of tuning to do. We've also been working on getting ready to use the scope to attain really robust time course measurements.
Outreach Summary
Made a lot of progress in planning and solidifying plans for education outreach. Alyssa has been in contact with Deanna Marroletti from Camp Launch and she will be visiting with her students on Wednesday, July 19th. We are planning on doing a DNA extraction activity, RFP streak, and gel electrophoresis. We finished the database up to 2008 prize-winning teams. We are now going to go to 2016 nominated teams.
Week 8 (July 17-July 23)
Wetlab Summary
Looking at analysis from FACS run, we are indeed seeing growth effect problems, and that's having an impact on fluorescence. We have working pTet pdts. We're geneblocking pLac mf-Lon.
Outreach Summary
We held the second event for our Bioengineering Speaker Series. About 30 people attended. We also have a potential collaboration going with Senator Mason. He wants to help us with outreach and invited us to a meeting. On Wednesday, we had a great time with two groups of rising 8th graders (16 students in each group). We did 2.5 hour lessons that incorporated strawberry DNA extraction, gel electrophoresis, and RFP streak. On Saturday, we did a presentation for Focusing on the Future, which was a career fair of sorts for the campers.
Week 9 (July 24-July 30)
Wetlab Summary
This week, we ran induction curves on J23100 pBad parts with pdts to try to get confirmation of functionality as well as to determine optimal concentrations. We also ran induction curves on working pTet combo parts but found fluorescence values to differ largely among constructs (could be attributed to bad luck with inoculating things.) We tried to run another time course to see if the time to steady state changes when a pdt + mf-Lon is added, but having weird issues with the FACS clogging--hopefully this can be solved by next week.
Outreach Summary
We held the third event for our Bioengineering Speaker Series. Dr. Puzey gave a fascinating presentation on how plants can have applications to bioengineering. Dr. Saha prepared delicious "plant-based, mind-altering" food for the audience.
Week 10 (July 31-August 4)
Wetlab Summary
This week, we ran induction curves on J23100 pBad parts with pdts to try to get confirmation of functionality as well as to determine optimal concentrations. We also ran induction curves on working pTet combo parts but found fluorescence values to differ largely among constructs (could be attributed to bad luck with inoculating things.) We tried to run another time course to see if the time to steady state changes when a pdt + mf-Lon is added, but having weird issues with the FACS clogging--hopefully this can be solved by next week.
Outreach Summary
Continued adding and editing entries for the database. Alyssa worked on the Bioengineering Lab website.
End of Summer
Week 11 (July 31-August 4)
Wetlab Summary
This week, we ran induction curves on J23100 pBad parts with pdts to try to get confirmation of functionality as well as to determine optimal concentrations. We also ran induction curves on working pTet combo parts but found fluorescence values to differ largely among constructs (could be attributed to bad luck with inoculating things.) We tried to run another time course to see if the time to steady state changes when a pdt + mf-Lon is added, but having weird issues with the FACS clogging--hopefully this can be solved by next week.
Outreach Summary
Continued adding and editing entries for the database. Alyssa worked on the Bioengineering Lab website.