Gibson Assembly
A modification of this method, Gibson assembly, described by Gibson et al. allows for single-step isothermal assembly of DNA with up to several hundreds kb. By using T5 exonuclease to 'chew back' complementary ends, an overlap of about 40bp can be created. The reaction takes place at 50°C, a temperature where the T5 exonuclease is unstable. After a short timestep it is degraded, the overlapps can anneal and be ligated. Cambridge University IGEM team made a video describing the process.
Read more about this topic: Polymerase Cycling Assembly
Famous quotes containing the words gibson and/or assembly:
“The designers [of the 1930s] were populists, you see; they were trying to give the public what it wanted. What the public wanted was the future.”
—William Gibson (b. 1948)
“A man may be a heretic in the truth; and if he believe things only because his pastor says so, or the assembly so determines, without knowing other reason, though his belief be true, yet the very truth he holds becomes his heresy.”
—John Milton (1608–1674)