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Team:WalthamHS BioHawks/Project/E-Notebook
From 2014hs.igem.org
E-Notebook
Our Journey-
September 24: This was the first meeting for the iGEM synthetic biology club for people who were interested in the project and who wanted to get an overview of the club.
October 8: We looked at iGEM projects from past years and learned about the jamboree.
November 13: In this meeting, we learned about the basics of synthetic biology and discussed possible ideas for our project.
January 28: We learned more about how synthetic biology works, as well as narrowed our options for a project. Students were assigned projects to research.
February 11: People presented research on their respective topics.
February 27: This was the official iGEM team’s first meeting. We narrowed our ideas down to four plausible projects.
March 4: Everyone voted on a project, based on a variety of criteria. After much debate, we decided on creating bacteria that could easily break down protein based stains in a safe and biodegradable fashion.
March 7: Our wiki group posted the summary of our project on our iGEM project.
March 11: The lab people began to grow bacteria after school, called BBa_J04500 (Part A) and BBa_J04650 (Part B), both of which were maintained on pSB1AK3 (a plasmid backbone). This backbone made them resistant against ampicillin and kanamycin antibiotics. These two samples of bacteria were placed on separate agar plates and were then incubated at 37 degrees ºC for 24 hours.
March 13: The lab people began to transfer the bacterial Parts A and B from the plates and placed them into separate culture tubes, both which were filled with 5ml LB broth. They were incubated for 37 degrees ºC for 24 hours.
March 17: The lab people continued on to step 3: digest and cut Parts A and B from their backbones using the restriction enzymes provided by NEB. The enzymes used were EcoR1, Xba1, Pst1, and Spe1. These parts were incubated at 37 degrees ºC for 30 minutes and then 80 degrees ºC for 20 minutes. We then ran the product through a gel, but unfortunately, the gel didn't show any results. We could only conclude that the digest did not cut the DNA correctly.
March 20: We decided to continue the process in order to practice lab techniques. The lab people then met to do step 4: ligation. This is where we had to assemble Part A, Part B, and the linearized plasmid backbone pSB1C3. We incubated the product, following the ligation, at 16 degrees ºC for 30 minutes and 80 degrees ºC for 20 minutes.
March 25: The lab people began to perform the transformation. NEB donated 10 beta strain competent cells which took in our ligated plasmid. These cells were grown on a chloramphenicol plate and, from the cells that survived on the plate, the ones that were red had the correct plasmid. Our control was transformed correctly proving that our process during the ligation and transformation process was accurate.
April 3: In this meeting, we decided what had to be done, as the practice was over, and assigned new positions so that the wiki and poster could be completed.
April 10: At this meeting we discussed research and wiki progress.
May 15: The lab people performed a digest of Part A, the synthesized subtilisin gene, the plasmid, and the control, and then ran a gel to check the digest. The gel showed that the digest was unsuccessful. The digest and gel were repeated and was still unsuccessful.
May 21: At this meeting we divided up the remaining tasks between the group members.
May 29: In this meeting we started to plan for the wiki, the presentation, the poster, and our final lab steps.
June 5: Together we planned the next steps of our lab procedure. The wiki people began to put together our website.
June 6: The lab people ran a gel of the digest performed on May 15th to double check results. Still no digest.
June 10: The lab people digested DNA from the distribution kit with restriction enzymes. They ran the gel to find out if the restriction digest worked. The enzymes did not digest the DNA. The lab people also tested subtilisin's ability to degrade skim milk. Drops of subtilisin were added to agar plates covered in milk, and the reaction was incubated for one hour. Subtilisin appeared to degrade the milk.
June 12: This was a meeting where our group decided to go through the wiki and edit everything to make sure it would be ready for the competition. We also began to piece together our presentation.
June 16: The lab people performed the second round of the subtilisin on milk experiment. Two additional plates were used as control: drops of water on milk and drops of E. coli on milk. Subtilisin showed milk degradation in comparison to water and E. coli.
June 18: All the team members met in the library to work on the wiki.
June 19: All the team members met in the library to work on the wiki. We made final touches on our wiki and worked on the poster and presentation.
June 20: All the team members met in the library to work on the poster and presentation.
