Genomics Core Facility, Stony Brook University

Hierarchical Structure

1.

Genomics Core Facility:

a.Director - Dr. Eli Hatchwell, MA MB BChir (Cantab) DPhil (Oxon) BA (OU), Associate Professor of Research Pathology
b.Introduction (see details below)

2.

Subdivisions:

a.Affymetrix Core Facility
b.DNA Sequencing Facility
c.Bioinformatics Services
d.Array-based Comparative Genomic Hybridization (aCGH) service for genome-wide copy number analysis (see details below)
e.Mutation screening service (see details below)
f.Multiplex ligation-dependent probe amplification (MLPA) for molecular-based, region specific copy number assays (see details below)
g.Pyrosequencing (see details below)
h.Consultation service (see details below)

Introduction to Genomics Core Facility

The newly created Genomics Core Facility is charged with the mission of facilitating all aspects of research related to genetics/genomics at Stony Brook University. Dr. Hatchwell is a Medical Geneticist and research scientist, who hails from the UK, via Cold Spring Harbor Laboratory. He is well versed in all modern methods of genetic/genomic analysis.

The facility aims to provide scientists at Stony Brook with the tools required to maximize their research. In the longer term, it is hoped that this facility will pave the way for Stony Brook to become a center of excellence in translational medicine, with particular emphasis on human genetics.

The following represent new methods/tools which are now available in our facility. Fees are being worked out and will be available shortly. We have an open door policy in our facility and welcome both customers for our services and collaborations. We are always interested in new projects!

Instrument Scheduling

Due to high demanding for some instruments or staff availability, users wishing to use BioPlex-200 System, Pyrosequencing, QIAxcel (formerly eGene), qPCR are required to make reservations. Please click here to make your reservation.

aCGH

We use a whole human genome tiling path BAC array for copy number analysis. This array has an effective resolution of 150kb and spans 93% of the human euchromatic genome. In addition to the basic tools/reagents, we have sophisticated bioinformatics tools at our disposal, as well as a large database of copy number variation in normals (currently over 800 individuals). We also offer genome wide copy number analysis using any of the major platforms, including NimbleGen and Agilent oligonucleotide based microarrays.

Larger Image
Fine tiling NimbleGen oligonucleotide array used to map chromosome 3 breakpoints with enough resolution to generate junction fragments by PCR (Jasmin Roohi, Cristina Montagna, David H. Tegay, Lance E. Palmer, John C. Pomeroy, Susan L. Christian, Norma Nowak and Eli Hatchwell. Disruption of Contactin 4 in 3 Subjects with Autism Spectrum Disorder. Journal of Medical Genetics (Rapid online publication - March 18th 2008).

QIAxcel System (Formerly known as eGene)

For gel electrophoresis of large numbers of standard DNA samples (e.g., PCR products), impractical using agarose gels, we offer capillary based electrophoresis on the QIAxcel System (formerly eGene system - Qiagen recently acquired eGene). This is a fully automated system for the identification, quantification and sizing of unmodified dsDNA. The system is able to run a 96 well plate in approximately 1.5 hours ( a further 30 minutes is required for analysis).

We accept a minimum of one plate for a run.

         

See: http://www1.qiagen.com/news/QIAxcel.aspx for further details.

Bio-Plex200

We offer the Bio-Plex 200 system for running assays based on Luminex bead technology, including the complete range of kits available from Millipore, Bio-Rad, Marligen, Panomics and many \others, whether protein or nucleic acid based. Please enquire for further details.

See: Bio-Rad Bio-Plex page for further details.

Mutation Screening Service Using Melt Analysis

There are many situations in which it is necessary to screen candidate genes in large numbers of individuals in order to define rare, pathological variants. It is often cost-prohibitive to perform sequencing for genes with many exons, when large cohorts are to be studied. We have implemented a method that utilizes melt analysis of heteroduplexes, using technology from Idaho Technology. The method is based on the dye LC-Green, which intercalates ds DNA at very high saturation levels, such that any premature melting that results from mismatches can be easily detected. The method allows for screening of exons in large sample sets at a fraction of the cost of sequencing. Any putative variants (often representing only a small percentage of the total) are validated by sequencing.

         

For more information on melt analysis, please visit http://www.idahotech.com/LightScanner/index.htm

Multiplex Ligation-Dependent Probe Amplification (MLPA)

While whole genome copy number detection using aCGH is very powerful, there are situations where it is necessary to target specific regions of the genome for copy number analysis, including:

a.Estimating the copy number of transgenes in animal models;
b.Validating interesting changes found on aCGH;
c.Focusing on specific regions of interest for screening large cohorts in candidate gene regions (sequencing/melt analysis does not detect deletions/duplications of genes);

MLPA is a very quantitative method for assessment of copy number - much more quantitative than qPCR (real-time PCR).

For more information on MLPA, please visit http://www.mrc-holland.com/pages/indexpag.html

Pyrosequencing (PyroMark MD machine)

Pyrosequencing is a highly quantitative sequencing method used in a variety of applications, including analysis of the methylome, detection of rare variants and population analysis.

For more information on Pyrosequencing, please visit http://www.pyrosequencing.com/

Consultation Service

We are available to aid in the design of assays and advise on the best tools/methods to use to answer any specific scientific question(s). We strongly advise users to discuss their project goals prior to embarking on the use of any given platform.