Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity
Type
Blog Post
Published
Oct. 11, 2018, 12:30 p.m.
... assembly reaction**
1
5 min., 37 ºC (2.5 μl)
687
100%
274,200
2,742,000
1
60 min., 37 ºC (2.5 μl)...expected complete sequence for the lacI/lacZ genes (1), while sequencing of white colonies showed a mixture...representative transformation plates obtained from 1-, 12- and 24-fragment assemblies of the lacI/lacZ ...lacZ cassette, and illustrates how the volume of the 1 ml outgrowth spread on each transformation plate can...fidelity studies conducted by Potapov et al. at NEB (1, 2). While T4 DNA Ligase, the mainstay of most biotechnological...over 90% for 24-fragment assemblies (Figure 3, Table 1). These improvements (5- to 12-fold increases in the... 16 ºC) x 30 (100 μl)
78
90.7%
783
9,792
Table 1. Yields and fidelities for Golden Gate Assemblies ...