Long-read sequencing for rare disease research workflows

If your rare disease research workflow stalls after short-read or tiered assays, HiFi long-read sequencing can add variant, methylation, and phasing evidence. In this PacBio webinar, Nina Gonzaludo, PhD, and Ramakrishnan (Ramki) Rajagopalan, PhD, from the Children's Hospital of Philadelphia, connect that workflow question to CHOP research cohort examples involving NIPBL insertions, a MEG3 deletion with methylation and phasing evidence, and Iso-Seq transcript evidence near HK1. Key takeaways Why rare disease research workflows can leave structural variants, repeat insertions, phasing relationships, methylation patterns, and transcript effects unresolved. What published rare disease studies show about HiFi WGS, including variant classes, methylation patterns, phasing, and workflow questions that are difficult to interpret with staged short-read and assay-specific approaches. How HiFi long-read sequencing supports research into SNVs, indels, CNVs/SVs, tandem repeats, methylation, and phasing in one data type. CHOP cohort examples involving NIPBL insertion events, a 203 bp MEG3 deletion phased with a nearby SNV, and an HK1 upstream duplication explored with PacBio Iso-Seq. Q&A on when to consider long reads earlier, mitochondrial DNA workflows, methylation tissue specificity, targeted versus genome-wide approaches, and 20x versus 30x coverage tradeoffs. Featured speakers Nina Gonzaludo, PhD, Global Segment Lead - Clinical, Rare Disease, PacBio Ramakrishnan (Ramki) Rajagopalan, PhD, Director of Translational Bioinformatics, Division of Genomic Diagnostics, Children's Hospital of Philadelphia Chapters 00:00 Welcome and webinar setup 01:33 Rare disease research workflows and why more complete data matters 02:49 HiFi sequencing overview for rare disease research 03:55 Rare disease study examples using HiFi whole genomes 09:50 Multiomic applications: Iso-Seq, methylation, and targeted sequencing 14:09 PacBio WGS variant pipeline and scale 17:11 Ramki Rajagopalan introduces CHOP and study goals 19:47 Clinical testing context and research workflow boundaries 20:30 Why tiered assays persist in practice 25:22 Gaps in staged rare disease research workflows 30:06 Small insertion and phasing in an unresolved sample 31:36 NIPBL insertions in Cornelia de Lange syndrome research 34:53 Multiomics examples from CHOP 37:22 MEG3 deletion, methylation, and Kagami-Ogata syndrome 40:23 Iso-Seq and HK1 transcript evidence 42:47 Barriers to bringing long reads toward lab implementation 45:07 Q&A: selecting samples for long-read sequencing 48:44 Q&A: multiomics, methylation, and RNA 50:00 Q&A: mitochondrial DNA and heteroplasmy 52:10 Q&A: methylation signatures and tissue specificity 53:02 Q&A: targeted versus genome-wide approaches 54:49 Q&A: coverage and variant calling tradeoffs 56:19 Q&A: downstream research follow-up and future progress Resources: HiFi sequencing technology: https://www.pacb.com/technology/hifi-... Rare disease HiFi sequencing whitepaper: https://www.pacb.com/wp-content/uploa... Long-read RNA-seq in rare disorders: https://www.pacb.com/publications/hif... Where would long-read data add the most value in your rare disease research workflow? Comment below. Subscribe to PacBio for more webinars on HiFi sequencing, rare disease research, human genomics, and multiomic applications:    / @pacificbiosciences   For Research Use Only. Not for use in diagnostic procedures. #RareDisease #HiFiSequencing #LongReadSequencing