Supplementary Figure 1: Screening strategy for CEL-HYB.

(a) Schematic outline of the long-range duplex PCR assay (one common forward primer, two reverse primers) developed to detect CEL-HYB. The allele was amplified by primers L11F and CELP-VNTR-R, which bind to CEL- and CELP-specific sequence, respectively. For the CEL-CELP wild-type allele, the PCR product that would be generated by these two primers is too long (19 kb) to be detected by the assay. Therefore, amplification of CEL-WT by the primers L11F and IAR was included as internal control. (b) PCR products from the assay visualized by agarose gel electrophoresis. The sizes of the CEL-WT- and CEL-HYB-specific amplification products are 4.2 kb and 3.2 kb, respectively. For CEL-HYB-negative samples, only the upper CEL-WT band is seen. For CEL-HYB-positive samples, two bands are detected. The CEL-WT product was designed to be larger than the CEL-HYB-specific product in order to verify the quality of the DNA sample. The gel picture shows screening of six samples (lanes 1-6). B, blank; N, negative control; P, positive control.