The GFP project has begun at BOSSLAB. Headed by the Chief Troublemaker of this blog (me), a group of people will be using BOSSLAB to play around with pGREEN, a plasmid that produces GFP.
The goal of this project is not only to get local people working hands-on with biotech, but to support the DIYBIO community by producing a model project, with documentation, that people can do. Over the next month or so I will be cranking out all the information you need to safely create and modify genetically engineered organisms in an environment about as sterile and complicated as your kitchen.
The GFP project will have # components, as listed below:
- Transform pGREEN intoe. Coli MM294
- Extract pGREEN from the transformants and verify the quality of the plasmid DNA
- Extract GFP from the transformants for extra points
This week was kind of like “Week 0” for the project in that I showed everyone how to pour plates, streak for individual colonies, and practice (some amount) of aseptic technique. As I mentioned, one of the goals for me in leading this project is to show people DIYBIO to show people basic techniques. To make sure I do a good job, I will be covering the techniques we use in the project in detail in separate posts.
Hopefully we will get to do the first item next weekend, at the twice-montly BOSSLAB meeting. It will be nice to have something really going on during the meeting; maybe more people will get interested and join the project, allowing us to get some nicer things for the project.
After this project comes ????. That means I don’t know if we will continue doing molecular biology, or switch to something like microbial diversity, or try to do some kind of Mendelian genetics project with plants. There are a lot of really cool biological things to investigate, and although molecular biology and synthetic biology are my favorites, BOSSLAB is really open to anything. If we continue in the molecular biology direction, we could try doing a restriction digest on the extracted plasmid and run it in a gel, or have part of it sequenced (find out what kind of GFP we are making!), or ligate it to a localization tag and see it expressed in only part of the cell (contingient on getting our fluorescence scope working). Really, there are all kinds of possibilities for this! But first, extraction and verification have to happen.
2 thoughts on “The GFP Project Week 0: Welcome to BOSSLAB”
Nice fluorescence! Please tell me which plasmid and expression cell line you used, and where I can get it from.
I used the pgreen plasmid and mm294 stain of e coli from carolina biological.