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:
“They wore the expression men always wore when they watched you dance, staring real hard but locked up inside themselves at the same time, so their eyes told you nothing at all and their faces, in spite of the sweat, might have been carved from something that only looked like flesh.”
—William Gibson (b. 1948)
“Our assembly being now formed not by ourselves but by the goodwill and sprightly imagination of our readers, we have nothing to do but to draw up the curtain ... and to discover our chief personage on the stage.”
—Sarah Fielding (1710–1768)