Team:Fudan-TSI/Experiments
Experiments
2019 Team Fudan-TSI works at the laboratory of Prof. Liang Cai in the School of Life Sciences of Fudan University. We mainly use the experimental devices and protocols within the laboratory, which are performed and optimized through years of practice.
Particularly, the fluorescence intensity and OD600 measurement and calibration protocols we use are from iGEM official measurement protocols.
Project by Team:Fudan-TSI
Mutation library generation is critical for biological and medical research, but current methods cannot mutate a specific sequence continuously without manual intervention. We hereby present a toolbox for in vivo continuous mutation library construction. First, the target DNA is transcribed into RNA. Next, our reverse transcriptase (RT) reverts RNA into cDNA, during which the target is randomly mutated by our RT's enhanced error-prone ability. Finally, the mutated version replaces the original sequence through recombination. These steps will be carried out iteratively, generating a random mutation library of the target with high efficiency as mutations accumulate along with bacterial growth. Our toolbox is orthogonal and provides a wide range of applications among various species. R-Evolution could mutate coding sequences and regulatory sequences, which enables the evolution of individual proteins or multiple targets at a time, promotes high-throughput research, and serves as a foundational advance to synthetic biology.