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. Author manuscript; available in PMC: 2019 Jun 10.
Published in final edited form as: Pharmacogenomics. 2010 Jan;11(1):43–53. doi: 10.2217/pgs.09.133

Figure 4. Further characterization of the tandem hybrid gene locus using long-range PCR.

Figure 4.

Fragment designations correspond to those in Figure 1. The genotypes of positive and negative control DNAs are as indicated. PCR product lengths are given in kb to the right, marker sizes on the left. All PCR reactions were carried out on genomic DNA. (A) Series of PCR reactions carried out with CYP2D8 forward and CYP2D6 reverse primers binding to intron 1, intron 6 and exon 9 sequences, demonstrate that each hybrid is located immediately downstream of CYP2D8. According to their respective hybrid composition, CYP2D6*77 produced all three amplicons, while CYP2D6*78 and CYP2D6*76 produced two and one, respectively. (B) In this series of reactions, the forward primer is CYP2D7 specific (also used for fragment H). As in (A), the hybrid tandems amplified fragments according to their hybrid structure. These shorter fragments readily amplify in less than 3 h and provide a convenient way to further analyze hybrids. Note that amplicon sizes vary slightly (middle and bottom panels) due to sequence differences between the hybrids.