S

Lane 1:

RhCcEe 1200bp RhD 600l>p

Rli.0 mbp KJiCVEe 129bp

Fig. 1. RHD genotyping: (A,B) Illustrations of targeted exons and introns of RHCE, RHD, and RHDy for RHD and RHD^genotyping, respectively (14,18,23,27). (C) RHD genotyping by ASPCR and 2% agarose gel electrophoresis. Products of each analysis are run in pairs; the intron 4/exon 10 standard RHD reaction is in the first lane of each pair and the RHD ^reaction is in the second lane of each pair. The RHD amplification generates 600-bp and 193-bp RHD-specific products, as well as 1200-bp and 129-bp internal control products derived from RHCE. The RHD^reaction generates a 381-bp product from a normal RHD allele and a 418-bp product from RHDy alleles, a primer pair targeting RhCE provides an internal control (14,18,23,27). Set A: normal RhD-negative individual; set B: normal RHD-positive individual; set C: normal RHD-positive/RHDy heterozygous individual; panel D/panel E: prenatal genotyping case involving a serologically RhD-negative/standard genotype-positive mother possessing the RHDy allele (panel D), and the initially ambiguous fetal genotyping (panel E). Lane S: 100-bp DNA ladder (Gibco/BRL, Gaithersburg, MD).

Fig. 1. RHD genotyping: (A,B) Illustrations of targeted exons and introns of RHCE, RHD, and RHDy for RHD and RHD^genotyping, respectively (14,18,23,27). (C) RHD genotyping by ASPCR and 2% agarose gel electrophoresis. Products of each analysis are run in pairs; the intron 4/exon 10 standard RHD reaction is in the first lane of each pair and the RHD ^reaction is in the second lane of each pair. The RHD amplification generates 600-bp and 193-bp RHD-specific products, as well as 1200-bp and 129-bp internal control products derived from RHCE. The RHD^reaction generates a 381-bp product from a normal RHD allele and a 418-bp product from RHDy alleles, a primer pair targeting RhCE provides an internal control (14,18,23,27). Set A: normal RhD-negative individual; set B: normal RHD-positive individual; set C: normal RHD-positive/RHDy heterozygous individual; panel D/panel E: prenatal genotyping case involving a serologically RhD-negative/standard genotype-positive mother possessing the RHDy allele (panel D), and the initially ambiguous fetal genotyping (panel E). Lane S: 100-bp DNA ladder (Gibco/BRL, Gaithersburg, MD).

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