Ye Bang-Ce, Chu Xiaohe, Fan Ye, Li Songyang, Yin Bincheng, Zuo Peng

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Presentation transcript:

Simultaneous Genotyping of DRB1/3/4/5 Loci by Oligonucleotide Microarray Ye Bang-Ce, Chu Xiaohe, Fan Ye, Li Songyang, Yin Bincheng, Zuo Peng The Journal of Molecular Diagnostics Volume 7, Issue 5, Pages 592-599 (November 2005) DOI: 10.1016/S1525-1578(10)60592-2 Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 A: The principle of PCR amplification and labeling of DRB gene, schematic representation of the exon 2. Figure indicates the relative positions of primers (the sequences were shown as underlined segments of the lowercase). The sequence of exon 2 is in gray. B: PCR products (289-bp) of exon 2 of DRB1/3/4/5 genes using the generic DRB primers (GH46 and RBAMP-B) were visualized by agarose gel electrophoresis. The Journal of Molecular Diagnostics 2005 7, 592-599DOI: (10.1016/S1525-1578(10)60592-2) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 The double-stranded PCR products and single-stranded products generated from linear PCR of the sample (IHW9382) were applied to the oligonucleotide array. The results demonstrated that single-stranded products generated much higher signals than double-stranded DNA. Subgroup A1: A01, A02, A03, A04, A04′, A05, A06, A08, and A12; subgroup A2: A07, A09, A10, and A11. White bars: The fluorescence signals of hybridization with single-stranded DNA. Gray bars: The fluorescence signals of hybridization with double-stranded DNA. The Journal of Molecular Diagnostics 2005 7, 592-599DOI: (10.1016/S1525-1578(10)60592-2) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 The amplicons of DRB loci of IHW9006 were hybridized with B01/B02 probes with different oligonucleotide length immobilized on slides. The B01/B02 probes of group B were selected to achieve significant discrimination of single-base mismatch, considering the strength and ratio of fluorescence signal of B01/B02 probe pair. The Journal of Molecular Diagnostics 2005 7, 592-599DOI: (10.1016/S1525-1578(10)60592-2) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 Procedure for analysis of hybridization data are illustrated for the sample (DRB1*04, DRB4). All oligonucleotide probes were spotted and named (refer to Table 2 for probes). After hybridization, signal intensities were quantified. The Journal of Molecular Diagnostics 2005 7, 592-599DOI: (10.1016/S1525-1578(10)60592-2) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 5 Hybridization results of the 32 IHW cell lines serving as reference samples. A gray box indicates a positive signal of hybridization, and a white box indicates a negative signal of hybridization. The Journal of Molecular Diagnostics 2005 7, 592-599DOI: (10.1016/S1525-1578(10)60592-2) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions