Detection of genome editing at the CCR5 locus by TALENs using HRMA.

<p>The gDNA of HEK293T cells mock-transfected, or transfected with pEGFP-N1 alone or with the left (L) or right (R) TALEN or both (L + R). The gDNA of transfected cells were subjected to qPCR using primers SK144 and SK145 and analyzed by HRMA post-amplification as described in Material and Methods. A) Normalized melt curves from indicated transfections. The y-axis shows normalized relative fluorescence units (RFU) while the x-axis depicts temperature (with a 0.2°C resolution between data points as detailed in Materials and Methods). Only 1°C temperature increments are shown for clarity. B) Difference curves were obtained by subtracting the normalized melting profile of each test sample from that of mock-transfected control. Two molecular clones were tested for each of the right and left TALENs. The pairs of TALENs used were designated as pair 1 or pair 2. sep: Separate transfections with either left or right TALEN alone.