Team:BostonU/Project Overview

From 2012.igem.org

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Latest revision as of 20:08, 25 October 2012

BostonU iGEM Team: Welcome

Project Overview

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:

Building: Build Genetic Circuits with MoClo Parts
Characterizing: Characterize Circuits using Flow Cytometry
Sharing: Generate Data Sheet for MoClo Parts

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

Title

Abandon All Hope, Ye Who PCR: MoClo and the Quest for Genetic Circuit Characterization

Authors

Monique De Freitas¹, Shawn Jin¹, Evan Appleton², Swati Carr², Sonya Iverson², Traci Haddock³, and Douglas Densmore^§

Affiliations

¹iGEM Team Member, ²iGEM Team Mentor, ³iGEM Team Advisor, ^§Faculty Sponsor, Department of Electrical and Computer Engineering, Boston University, Boston, MA, USA

Abstract

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.

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-One of our major goals this year is to introduce MoClo as an alternative assembly method for iGEM teams to use. Our other major goal is to generate a standard protocol for the characterization of genetic circuits containing fluorescent proteins and share this protocol with the synthetic biology community.
+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.
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-In order to achieve these goals, there are many steps that must be taken. First, we must convert BioBrick Parts into MoClo Parts using PCR strategies. Once the basic Parts are converted, our project will have three thrusts:
+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:
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-<p dir="ltr">Our project aims to introduce a standardized protocol for the characterization of genetic circuits using flow cytometry. We built a vast number of both simple and complex genetic circuits that were characterized using flow cytometry. These genetic circuits were built using an assembly technique called MoClo (developed by <a href="http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0016765"> Weber et al, 2011</a>), 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 BioBrick Parts from the <a href="http://partsregistry.org/Main_Page">Registry</a> into MoClo Parts using PCR and cloning strategies. We built and characterized various genetic circuits using MoClo Parts and compared them against their pre-existing BioBrick counterparts in order to compare the characterization results from the two assembly techniques. We also created a standardized data sheet 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.
+<p dir="ltr">
+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 <a href="http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0016765"> Weber et al., 2011</a>, 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 <a href="http://partsregistry.org/Main_Page">Registry</a> 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.
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