Format

Send to

Choose Destination
Nucleic Acids Res. 2014 Nov 10;42(20):e154. doi: 10.1093/nar/gku829. Epub 2014 Sep 8.

PaperClip: rapid multi-part DNA assembly from existing libraries.

Author information

1
School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3JR, UK School of Engineering, University of Edinburgh, Edinburgh, EH9 3JL, UK M.Trubitsyna@ed.ac.uk.
2
School of Biological Sciences, Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, EH9 3JR, UK.
3
School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3JR, UK.
4
School of Engineering, University of Edinburgh, Edinburgh, EH9 3JL, UK.
5
School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3JR, UK M.Trubitsyna@ed.ac.uk.

Abstract

Assembly of DNA 'parts' to create larger constructs is an essential enabling technique for bioengineering and synthetic biology. Here we describe a simple method, PaperClip, which allows flexible assembly of multiple DNA parts from currently existing libraries cloned in any vector. No restriction enzymes, mutagenesis of internal restriction sites, or reamplification to add end homology are required. Order of assembly is directed by double stranded oligonucleotides-'Clips'. Clips are formed by ligation of pairs of oligonucleotides corresponding to the ends of each part. PaperClip assembly can be performed by polymerase chain reaction or by cell extract-mediated recombination. Once multi-use Clips have been prepared, assembly of at least six DNA parts in any order can be accomplished with high efficiency within several hours.

PMID:
25200084
PMCID:
PMC4227759
DOI:
10.1093/nar/gku829
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Silverchair Information Systems Icon for PubMed Central
Loading ...
Support Center