PubReading [333] - Enzymatic assembly of DNA molecules up to several hundred kilobases - D. Gibson, H. Smith et al.
PubReading
English - May 31, 2023 11:40 - 10 minutes - 9.67 MBLife Sciences Science Health & Fitness Medicine Homepage Download Apple Podcasts Google Podcasts Overcast Castro Pocket Casts RSS feed
We describe an isothermal, single-reaction method for assembling multiple overlapping DNA molecules by the concerted action of a 5' exonuclease, a DNA polymerase and a DNA ligase. First we recessed DNA fragments, yielding single-stranded DNA overhangs that specifically annealed, and then covalently joined them. This assembly method can be used to seamlessly construct synthetic and natural genes, genetic pathways and entire genomes, and could be a useful molecular engineering tool.DOI:10.1038/NMETH.1318 - 2009