For DNA extraction, tail tissue was heated in 200 μl of chelex reagent (10% chelex, 0.1% Tween 20, and 1% proteinase K) for over 3 hours at 65°C, followed by proteinase K inactivation for 15 min at 75°C. For the polymerase chain reaction (PCR) reaction, 4 μl of the supernatant was placed into 16 μl of reaction mixture consisting of the AccuPower Taq PCR Master Mix (K-2609; Bionner) and forward and reverse primers (Macrogen). The DNA of cng3, abca4, and pde6b genes was amplified by PCR, and WT and mutant bands were detected as previously described (48, 49).

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