Figure 3
From: STAR: a simple TAL effector assembly reaction using isothermal assembly
Full-length 16mer TALEs in destination vectors are generated in a second Gibson reaction.
(a) Schematic of fully assembled TALE into destination vector showing both the primer binding sites for colony PCR and XhoI/NheI restriction sites for digest of TALE-TFs. (b) Colony PCR of bacterial colonies expressing a typical TALE-TF reaction. Clones expressing the fully assembled TALE-TF show amplicons of about 1.7 kb length (↓– clone potentially expressing full-length DNA binding domain; * – clones shown in panel c). (c) Clones identified in colony PCR are further characterised by restriction digest yielding a 2.4 kb fragment for fully assembled TALE-TFs (* – sequence-verified clones expressing full-length TALE-TF).
