Our project uses 3 major new technologies: Multiplex Automated Genome Engineering (MAGE), chip-based synthesis of DNA, and lamba red recombination, along with more traditional bioinformatics. We hope that future iGEM teams will also use these techniques in their own synthetic biology projects.

Zinc Finger Binding Site Finder

Check out our Zinc Finger Binding Site Finder Tool here! This tool was used to search the human genome for the six target DNA sequences that we used to design our custom zinc finger arrays.

Lambda Red- Mediated Recombineering

Useful Links

• Gene Knockouts and Exchanges by Linear Transformation: http://rothlab.ucdavis.edu/protocols/Lin.Transform.html
• Open Wet Ware Protocol: http://openwetware.org/wiki/Recombineering/Lambda_red-mediated_gene_replacement

Gibson (Isothermal) Assembly

Summary (Adapted from Gibson et al)^4,5

• Source: http://www.nature.com/nmeth/journal/v6/n5/full/nmeth.1318.html
• Protocol: http://www.nature.com/protocolexchange/protocols/554#/main

References

4. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA 3rd, Smith HO. (2009). Enzymatic assembly of DNA molecules up to several hundred kilobases. Nature Methods,6(5):343-5. Epub 2009 Apr 12. [http://www.nature.com/nmeth/journal/v6/n5/full/nmeth.1318.html]

5. Gibson D. (2009). One-step enzymatic assembly of DNA molecules up to several hundred kilobases in size. Nature Protocols,Published online 16 April 2009, doi:10.1038/nprot.2009.77. [http://dx.doi.org/10.1038/nprot.2009.77]