Core B - Virogenomics & Biostatistics Core

NIH RePORTER · NIH · P01 · $246,815 · view on reporter.nih.gov ↗

Abstract

Core B Project Summary/Abstract CORE B adheres to good clinical laboratory practices (GCLP) and operates under BSL-2+ containment. Hence, CORE B is able to work with infectious agents and infectious samples, including HIV positive samples and those that require worker immunization and/or federal clearance. Any clinical material used in projects 2, 3, 4, 5 will be processed in CORE B as well as tissue from EBV-infected humanized mice for Project 2. CORE B supports all projects of this program by providing NextGen sequencing, informatics and statistical services. (1) Real-time polymerase chain reaction (qPCR) is the standard for clinical quantitation of infectious agents such as HIV and herpesviruse. It is the method of choice to confirm microarray data or data obtained from RNAseq experiments. We developed real-time QPCR arrays for all human herpesviruses, for targeted profiling of cell signaling pathways such as NFκB target genes and for human microRNAs. These will be used in projects 2, 3, 4, and 5. (2) To facilitate NextGen sequencing the core use the Agilent Bioanalyzer for QC of input RNA and post qPCR analysis, and Tecan general pipetting robot for making libraries of any chemistry (Illumina, PacBio, 454, IonTorrent). CORE-B also operates a 454/GsJunior NextGen sequencer with 800bp read-length and a PGM/IonTorrent machine with 400bp read-length. The longer read-length capability has proven essential for sequencing across viral repeat regions in BAC mutants (projects 2, 3, 4, and 5) and HLA haplotyping (project 3). (2) Bioinformatics. CORE B supports a full time programmer. It maintains 100 TB of storage (~10,000 human genomes) and dedicated Linux (128GB RAM) and OsX (64GB RAM) servers, each with open source and commercial software to support DNAseq, RNAseq and MS/MS protein analyses. Furthermore, it serves as the conduit to access the wider UNC computing infrastructure and if need be Amazon EC cloud. (3) Biostatisticis. The core support a full time statistician (MS level and above) and Dr. Steven Marron, the Amos Hawley Distinguished Professor in Statistics, as a Co-Investigator, who will provide in depth statistical advice on experimental design and analysis as it pertains to cell culture, mouse and studies on patient material, such as proposed in projects 3, 4, and 5. We decided on using R as the analyses program of choice, but have also access to SAS (via a UNC site-license) and Phyton programming environments.

Key facts

NIH application ID
9986669
Project number
5P01CA019014-41
Recipient
UNIV OF NORTH CAROLINA CHAPEL HILL
Principal Investigator
Dirk P Dittmer
Activity code
P01
Funding institute
NIH
Fiscal year
2020
Award amount
$246,815
Award type
5
Project period
— → 2022-06-30