PubReading [333] - Enzymatic assembly of DNA molecules up to several hundred kilobases - D. Gibson, H. Smith et al.

<p>We describe an <strong>isothermal</strong>, single-reaction method for assembling multiple overlapping DNA molecules by the concerted action of a 5' <strong>exonuclease</strong>, a DNA <strong>polymerase</strong> and a <strong>DNA ligase</strong>. First we recessed DNA fragments, yielding single-stranded DNA overhangs that specifically annealed, and then covalently joined them. This <strong>assembly</strong> method can be used to seamlessly construct synthetic and natural genes, genetic pathways and entire genomes, and could be a useful molecular engineering tool.<em>DOI:10.1038/NMETH.1318 - 2009</em></p>