Title
Automated assembly of standard biological parts
Date Issued
01 January 2011
Access level
metadata only access
Resource Type
book part
Author(s)
University of California
Publisher(s)
Academic Press Inc.
Abstract
The primary bottleneck in synthetic biology research today is the construction of physical DNAs, a process that is often expensive, time-consuming, and riddled with cloning difficulties associated with the uniqueness of each DNA sequence. We have developed a series of biological and computational tools that lower existing barriers to automation and scaling to enable affordable, fast, and accurate construction of large DNA sets. Here we provide detailed protocols for high-throughput, automated assembly of BglBrick standard biological parts using iterative 2ab reactions. We have implemented these protocols on a minimal hardware platform consisting of a Biomek 3000 liquid handling robot, a benchtop centrifuge and a plate thermocycler, with additional support from a software tool called AssemblyManager. This methodology enables parallel assembly of several hundred large error-free DNAs with a 96+% success rate. © 2011 Elsevier Inc. All rights reserved.
Start page
363
End page
397
Volume
498
Language
English
OCDE Knowledge area
Biología celular, Microbiología
Subjects
Scopus EID
2-s2.0-79955706591
Source
Methods in Enzymology
ISSN of the container
00766879
Sponsor(s)
We thank Martin Pollard of the Joint Genome Institute ( www.jgi.doe.gov ) for helpful discussions on assembly line automation design. We also thank Nina Revko and the 2009 Berkeley computational iGEM team for early work on the automated assembly software. This work was funded by SynBERC ( www.synberc.org ).
Sources of information:
Directorio de Producción Científica
Scopus