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Figure 2. Oligonucleotide primers used to amplify ENAM coding exons and determine their DNA sequence. On the upper right is a 1% agarose gel, stained with ethidium bromide, showing each PCR amplification product that was used to identify the enamelin gene mutations. Above each band is the number of the ENAM exon that was amplified; below each band is the predicted length of the PCR amplification product. The exon-specific PCR amplification primer pairs (F = forward; R = reverse) and their annealing temperatures (T°C) are listed. The reactions had a five-minute denaturation at 94°C, followed by 40–50 cycles each with denaturation at 94°C for 30 sec, primer annealing at 55–61°C for 30 sec, and product extension at 72°C for 30–60 sec. In the final cycle, the 72°C extension was for 7 min.
