Chimeric proteins used in variants of this technique include the HhH domain of topoisomerase V from M. kandleri to reduce contamination. Early versions of this technique (10[1])had people manually swap tubes between water tubs (10[1])by (10[1])hand for (10[1])hours at a time. (10[1]-5[1])This (10[1])technique is used to generate libraries for MPRA (10[1])and deep mutational scanning experiments. KAPA (10[1])(“kappa”), Phusion, and Q5 are high-fidelity (10[1])enzymes used in this technique, which can be made intentionally defective by (10[1])replacing magnesium ions (10[1])with manganese. Betaine or DMSO can be added to samples in this technique with high GC contents. Troubleshooting steps (10[1])for this technique include temperature gradients (10[1])and adjusting the master mix’s (10[1])primer (10[1])concentration. (10[2])For 10 (10[1])points, name this technique consisting of denaturation, (10[1])annealing, and (10[1])extension phases that is used to amplify strands of DNA. ■END■ (10[1])