U of L Sequencing Technology Center
We offer single molecule, real-time (SMRT) sequencing on the Revio and Sequel II systems. SMRT sequencing is characterized by long read lengths and high intramolecular consensus accuracy, and can be used to sequence libraries ranging from 250bp – 50kb. Common applications include whole genome sequencing, full length isoform profiling, highly accurate amplicon sequencing, and Sanger sequencing replacement. Custom preparations and targeted sequencing approaches may be available upon request; please reach out via the contact form below to discuss further!
HiFi Whole Genome Sequencing
Library preparation and sequencing protocol that generate of 15-20 kilobase (kb) reads with >99.9% accuracy. Use these reads for de novo assembly of reference-grade genomes of model and non-model organisms. Currently enabled on the Revio system.
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Input requirements: 5-10 μg HMW gDNA in fragments >25kb
Low Input Whole Genome Sequencing
HiFi WGS libraries made with as low as 400ng genomic DNA. Used for extremely limiting or single organism samples. The resulting HiFi reads can be used for de novo assembly and variant analysis. Currently enabled on the Revio system.
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Input requirements: 400 ng gDNA, >20kb. Ultra-low protocols also available for >25ng gDNA; contact team for details.
Isoform Sequencing (IsoSeq)
Preparation and sequencing methods to resolve full-length mRNA transcripts using HiFi reads. Data often used for the generation of novel reference transcriptomes, and defining splicing patterns and/or alternate isoforms usage across experimental conditions. Currently enabled on the Sequel IIe system.
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Input requirements: at least 500ng total RNA with RIN > 7.
Amplicon Sequencing
Short and long-range amplicons can be sequenced on the Revio (> 5kb) or Sequel IIe (250bp - 5kb) systems with high accuracy (>99.9%), allowing for phasing of variants across the amplicon length, providing haplotyping capabilities within a complex mixture.
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Input requirements: Varied based on amplicon size and samples per pool; contact team to discuss details.
MAS-IsoSeq (single cell)
Preparation and sequencing of concatenated cDNA molecules containing cell- and mRNA-specific barcodes, developed in conjunction with 10X genomics. Provides resolution of isoform variation at the single cell level. Currently enabled on the Revio system
Input requirements: unfragemented cDNA following 10X cDNA generation; contact team for details.
Human Leukocyte Antigen (HLA) Sequencing
HLA profiling allows investigators to assess the association of HLA alleles and resilience/susceptibility to disease. Our team offers profiling of up to 11 class I and/or class II HLA genes, with up to 96 subject samples profiled per run. Enabled on the Sequel IIe system.
Input requirements: 500-1000 ng gDNA from each sample, dependent on loci screened