Genotyping of the G616A Substitution in the <i>TENP</i> Gene by the PCR-RFLP Method and Comparison of the Genotypes with <i>G2</i>^<i>A</i> and <i>G2</i>^<i>B</i> alleles.

<p>(A) Schematic representation of the <i>Hph</i>I-RFLP in the 506-bp PCR product amplified from genomic DNA (encompassing exon 7 and exon 8 of the <i>TENP</i> gene). Digestion with <i>Hph</i>I (5′-GGTGA(N)₈↓-3′ or 3′-CCACT(N)₇↑-5′) is predicted to produce three DNA fragments (166 bp, 63 bp, and 277 bp) in the <i>G2</i>^<i>A</i> allele, whereas two fragments are predicted (166 bp and 340 bp) from the <i>G2</i>^<i>B</i> allele. (B) Genotyping using 2% agarose gel electrophoresis of the PCR products digested with <i>Hph</i>I. The nucleotides at position 616 and genotypes of ovoglobulin G2 are shown at the bottom of the lanes. Lane 1‒2, A/A (<i>G2</i>^<i>A</i>/<i>G2</i>^<i>A</i>) in Ehime-Jidori; lane 3‒4, A/G (<i>G2</i>^<i>A</i>/<i>G2</i>^<i>B</i>) in Ehime-Jidori; lane 5‒6, G/G (<i>G2</i>^<i>B</i>/<i>G2</i>^<i>B</i>) in Ehime-Jidori; P, undigested PCR product; M, 100 bp DNA Ladder RTU (0.1‒3 kb) (NIPPON Genetics, Tokyo, Japan).