PaperClip: rapid multi-part DNA assembly from existing libraries

摘要: 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 multiple parts from currently existing libraries cloned in any vector. No restriction enzymes, mutagenesis internal sites, or reamplification add end homology are required. Order directed by double stranded oligonucleotides‐‐‘Clips’. Clips formed ligation pairs oligonucleotides corresponding the ends each part. PaperClip can be performed polymerase chain reaction cell extract-mediated recombination. Once multi-use have been prepared, at least six order accomplished with high efficiency within several hours.