Find the notebooks regarding our subgroups below. They show our daily work in chronological order. Summarized strategies, plans and results are found in the project or the HEARTBEAT section respectively. Note that we used abbreviations especially for plasmids and oligos. Find a more detailed description of those in the material section.
In this subproject we cloned natural promoters from a variety of sources (e. g. other research groups). We chose promoters that are somewhat specifically inducible. We then tested conditions that we found in literature to induce those promoters. Some of those experiments helped us to determine the induction conditions for our own synthetic promoters.
This was one of our largest subprojects. During the first few weeks of this year's iGEM we mostly worked on DNA synthesis and cloning of those promoters. Later on, the focus shifted to screening and characterizing those promoters as described in the methods and measurement sections.
We started out with getting used to cell culture work and learned the basics such as splitting, freezing and transfecting cells. For the latter, we started out by transfecting our fluorescent protein templates. Later on, the cell culture work became essential for any promoter testing we carried out.
As selecting stables is a very time consuming task, we tried to start as early as possible with this subproject. After several weeks of selection, cell lines with stably integrated FRT sites were established. After that, our focus shifted towards characterizing those cell lines by identifying the number and genomic localizations of those integration sites.
Removing restriction sites that are used for the BBb standard (such as PstI, EcoRI and other) from the fluorescent proteins was the first requirement to make those proteins usable for BioBricking. Later on, we created several standardized fluorescent proteins and localization tags. On the way, we tested the creation of fusion plasmids by using the BBb standard. Towards the end of our project, a lot of cloning work for the part submission had to be done.
The notebook of the HEARTBEAT (Heidelberg Artificial Transcription Factor Binding Sites Assembly and Engineering Tool) project comprises the work on three sublanes: HEARTBEAT database (DB), HEARTBEAT graphical user interface (GUI) and HEARTBEAT fuzzy modeling (FN) as well as some additional work on logo as well as wiki design.
The section consists of RT-PCR, flow cytometry, dual plasmid and microscopy notebook, summarizing our large variety of measurements.