In this video, you will get a brief outline of how you can go from a DNA sequence design to an actual plasmid which you can transform for your iGEM project. This includes going through some major molecular biology lab techniques.

After you have designed your plasmid with the appropriate parts, origin of replication and antibiotic marker, it’s time to take what you’ve designed on the computer to an actual molecule that you can transform! This DNA assembly can be done by repeating four techniques:

1. Restriction digests or cutting DNA. In this process, you cut DNA strands at specific sequences (called restriction sites) using proteins known as restriction enzymes. In a
biobrick plasmid, there are very specific restriction sites on both sides of your gene of interest. By using specific restriction enzymes, you can cut out the gene of interest from its original plasmid backbone.

2. Gel extraction or separating DNA strands. Before you can glue a strand of DNA into another plasmid backbone, you need to first separate your gene that you cut out from its original plasmid backbone. You can do this by using a technique called gel electrophoresis. In this technique, you use agarose gel to separate DNA based on its size.

3. Ligation or gluing different strands of DNA. After extracting your appropriate DNA strands, you can glue together the different pieces of DNA you want to make your device. This can be done by using an enzyme called DNA ligase.

4. Transforming or adding the plasmid to the cells. After ligating your gene of interest into the plasmid backbone, you can transform this circular plasmid into your cell. The cells can replicate and make more of your DNA. In order to get back your plasmid, you
can culture your cells (making sure that you use the correct antibiotic so that only the cells that have your plasmid grow), and miniprep or extract the plasmid from the cells.

Repeating these four steps, you can start to build your genetic device from individual genes in the registry or those of your making. Now that you have an idea of all the tools needed for a synthetic biology project, you can start thinking more about forming an iGEM team and what that constitutes! :)