The introduction of new DNA sequencing technologies presents an exceptional opportunity for creative applications with potential for breakthrough discoveries. Funding is requested for a Pacific Biosystems (PacBio) sequencing instrument. The newly available PacBio third generation sequencing technology will be located in the Cornell University Life Sciences Core Laboratories Center (CLC). The CLC has extensive experience in running next generation sequencing instrument systems and in providing a wide range of next generation sequencing applications as core facility services. The CLC currently has a Roche 454 GS FLX, two Illumina GAIIx units and an Illumina HiSeq 2000 sequencing platform. There is a clear need for a PacBio instrument to be placed at Cornell. This application describes 24 projects from 21 NIH funded investigator groups that will utilize the instrument. The PacBio will be operated with a multidisciplinary, multi-core approach, including support from the CLC advanced technology assessment, genomics, epigenomics, bioinformatics, and bio-IT core resources. Implementation of the PacBio in a core facility as a university-wide shared research resource will enable cost effective access to this vital new technology. The PacBio instrument system allows single molecule real time sequencing. The system includes high throughput sample preparation, sequence detection, data processing, and assembly. There is no other instrument currently available that can sequence with a comparable combination of PacBio long read lengths, fast throughput, and low cost per run. PacBio applications include de novo assembly of smaller scale genomes, hybrid de novo and resequencing assembly of larger genomes that cannot be solved using short- read technologies alone, rare mutation (SNP) detection, characterization of structural variation, and gene expression and whole transcripts sequencing using cDNA libraries. PacBio applications under development include direct detection of DNA methylation by measuring polymerase kinetics, direct RNA sequencing, direct detection of RNA modifications such as RNA methylation, and high-resolution studies of macromolecular complexes like the ribosome and the spliceosome,. These applications are unique features of the PacBio technology. The PacBio technology will make possible experiments that are impractical or impossible with other available sequencing instruments. The instrument will facilitate a broad range of current and future NIH funded projects at Cornell University.
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