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Team:BostonU/Project Overview
From 2012.igem.org
Project Overview
- Building: Build Genetic Circuits with MoClo Parts
- Characterizing: Characterize Circuits using Flow Cytometry
- Sharing: Generate Data Sheet for MoClo Parts
Our project has two major goals: 1. To introduce MoClo as an alternative assembly method for use by iGEM teams and 2. To develop a standard protocol for the characterization of genetic circuits containing fluorescent proteins and share this protocol with the synthetic biology community.
In order to achieve these goals, there many steps must be taken. First, we must convert BioBrick Parts into MoClo Parts using PCR. Upon converting basic parts, our project will have three thrusts:
As we worked towards our first goal over the summer months, we ran into difficulties with some of our PCR and cloning reactions. This has unfortunately delayed our other goals, but we're still working hard towards generating a characterization workflow and a MoClo data sheet format for the Jamboree.
Abstract
Abandon All Hope, Ye Who PCR: MoClo and the Quest for Genetic Circuit Characterization
Monique De Freitas¹, Shawn Jin¹, Evan Appleton², Swati Carr², Sonya Iverson², Traci Haddock³, and Douglas Densmore§
¹iGEM Team Member, ²iGEM Team Mentor, ³iGEM Team Advisor, §Faculty Sponsor, Department of Electrical and Computer Engineering, Boston University, Boston, MA, USA
Our project has three aims: to introduce MoClo as an alternative assembly technique to BioBricks, to develop a standardized protocol for the characterization of genetic circuits using flow cytometry, and to share our MoClo Kit with the iGEM community. MoClo is an assembly technique developed by Weber et al., 2011, which involves a multi-way, one-pot digestion-ligation reaction, enabling faster and more efficient construction of genetic circuits. We converted a large subset of BioBricks from the Registry into MoClo Parts using PCR and cloning strategies. We are in the process of building and characterizing various genetic circuits using MoClo Parts, which we will compare to their BioBrick counterparts. A characterization workflow will be shared once this is complete. We also developed a data sheet using Clotho to be included in the Registry of Standard Biological Parts for each Part we characterized to easily share our data with the synthetic biology community.
