In this section
@article{Casini:2013:nar/gkt915,
author = {Casini, A and MacDonald, JT and De, Jonghe J and Christodoulou, G and Freemont, PS and Baldwin, GS and Ellis, T},
doi = {nar/gkt915},
journal = {Nucleic Acids Research},
title = {One-pot DNA construction for synthetic biology: the Modular Overlap-Directed Assembly with Linkers (MODAL) strategy},
url = {http://dx.doi.org/10.1093/nar/gkt915},
volume = {42},
year = {2013}
}
TY - JOUR
AB - Overlap-directed DNA assembly methods allowmultiple DNA parts to be assembled together inone reaction. These methods, which rely onsequence homology between the ends of DNAparts, have become widely adopted in syntheticbiology, despite being incompatible with a key principleof engineering: modularity. To answer this, wepresent MODAL: a Modular Overlap-DirectedAssembly with Linkers strategy that brings modularityto overlap-directed methods, allowing assemblyof an initial set of DNA parts into a variety ofarrangements in one-pot reactions. MODAL isaccompanied by a custom software tool thatdesigns overlap linkers to guide assembly,allowing parts to be assembled in any specifiedorder and orientation. The in silico design of syntheticorthogonal overlapping junctions allows formuch greater efficiency in DNA assembly for avariety of different methods compared with usingnon-designed sequence. In tests with three differentassembly technologies, the MODAL strategy givesassembly of both yeast and bacterial plasmids,composed of up to five DNA parts in the kilobaserange with efficiencies of between 75 and 100%.It also seamlessly allows mutagenesis to beperformed on any specified DNA parts duringthe process, allowing the one-step creation of constructlibraries valuable for synthetic biologyapplications.
AU - Casini,A
AU - MacDonald,JT
AU - De,Jonghe J
AU - Christodoulou,G
AU - Freemont,PS
AU - Baldwin,GS
AU - Ellis,T
DO - nar/gkt915
PY - 2013///
SN - 1362-4962
TI - One-pot DNA construction for synthetic biology: the Modular Overlap-Directed Assembly with Linkers (MODAL) strategy
T2 - Nucleic Acids Research
UR - http://dx.doi.org/10.1093/nar/gkt915
UR - http://hdl.handle.net/10044/1/26424
VL - 42
ER -
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