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Fully phased, allele-specific HLA sequencing – the perfect pair. - 2 Pages

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Fully phased, allele-specific HLA sequencing – the perfect pair.
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Catalogue excerpts

PACBIO* Single Molecule Real-Time (SMRT®) Sequencing delivers reads that span the lengths of the majority of HLA class I and II genes. Unambiguously phased 4-field HLA types without imputation. With a more accurate and complete picture, gain deeper understanding of immune-related disease causality, graft-versus-host disease in hematopoietic transplantation, and drug hypersensitivity1. - Phase polymorphisms across SNP-poor regions - Achieve allele-level segregation - Detect variants in regulatory regions within 5' UTRs, introns, and 3' UTRs - Characterize full-length transcribed minor variants of HLA alleles - Obtain direct evidence for new HLA alleles through de novo, reference-free consensus generation - Flexibly adjust amplicon size (e.g., phase through either exons 2, 3 and/or 4, or the entire HLA gene) or scale project size with cost-effective multiplexing solutions VALIDATION OF FULL-LENGTH HLA CLASS I AND EXON 2-4 LONG AMPLICON HLA CLASS II SEQUENCING ACROSS 96 SAMPLES No of Expected Alleles I Alleles Discordant I Discordant Alleles    I Non-validated I to Pre-types    I Orthogonally Validated I    Discordant SMRT Sequencing of 96 samples correctly identified 863 unique allele types and 22 potentially discordant alleles compared to low-res UCLA pre-type data. Orthogonal validation with Sanger Sequencing (of the 18 samples tested) demonstrated 100% of the potential discordant alleles matched the PacBio data. Four remaining alleles have not been validated and one DQB1 allele was miscalled due to PCR-related allele-drop out3.

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FROM TARGETED REGION TO FULLY PHASED, ALLELE-SPECIFIC HLA TYPES Locus specific HLA primer Library Preparation -- Utilize off-the-shelf enrichment methods or develop your own assays Customer provided assays Commercial primers -- Multiplex amplicons that span partial or full-length HLA genes -- Support available for library automation SMRT Sequencing with PacBio® Systems -- Span the majority of HLA class I and II genes with ~10 kb average read lengths -- Sequence HLA class I and II genes in 2-3 hours -- Identify > 16K full-length alleles per week PacBio RS II System -- Achieve consensus accuracies...

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