A simple method for site-directed mutagenesis using the polymerase chain reaction.
Tadege, M. et al. Large-scale insertional mutagenesis using the Tnt1 retrotransposon in the model legume Medicago truncatula.
Mutations A160T, A205P, and D266G were created by site-directed mutagenesis using the QuickChange site-directed mutagenesis kit (Agilent).
GFP-BFL1ΔC (lacking the C-terminal 24 residues) was generated by PCR-based mutagenesis using the Quickchange mutagenesis kit (Stratagene).
Mutant DNAs were generated by PCR-based mutagenesis using the Quickchange mutagenesis kit (Stratagene), or were purchased at GenTech.
Targeted gene mutagenesis using the CRISPR/Cas9 system has been achieved in several orders of insects.
An additional unannotated SalI restriction site in the pGreen0229 backbone was removed by PCR-based silent mutagenesis using the NOS-mut_for1/NOS_mut_rev1 primer pair.
Four different mutations (V157F, R175H, R249S, and R273H) were generated by site-directed mutagenesis using the QuikChange II XL site-directed mutagenesis kit (Agilent Technologies).
The L834R-EGFR pCDNA3 and T766M-EGFR pCDNA3 plasmids were produced through site-directed mutagenesis using the primers described in Supplementary Table 5.
Reetz, M.T., Soni, P., Fernandez, L., Gumulya, Y. & Carballeira, J.D. Increasing the stability of an enzyme toward hostile organic solvents by directed evolution based on iterative saturation mutagenesis using the B-FIT method.
GFP-tagged constructs were made by cloning CSA WT or CSA 8M, which was created by site-directed mutagenesis using the QuickChange site-directed mutagenesis kit (Agilent), into pENTR1A-GFP-N2 (Addgene).
