The Nucleic Acid Shared Resource (NASR) provides researchers with a variety of instrumentation platforms and expertise for DNA sequencing, genotyping, DNA methylation analysis and quantitative real-time PCR. It also offers accessory equipment for quantitative measurement and quality control of nucleic acids, and nucleic acid imaging.
Sanger-Based Sequencing and Genotyping Equipment
The instrument platform is the Applied Biosystems 3730 DNA Analyzer, with 48 capillary arrays, automated loading and a 16x96-well plate stacker for loading. Data collection v3.0 is supplied with the instrument, which manages instrument setup, controls instrument operations, allows real-time data visualization and performs diagnostics. Sequencing Analysis v5.2 and GeneMapper® v3.7 are also included.
- Instrument 1 Specifications: 50cm array dedicated to sequencing, long run (4 hrs): 800-1100 bp
- Instrument 2 Specifications: 36cm array dedicated to short fragments. Genotyping of STRs (1 hr): Size Range 80 – 489 bp, genotyping of SNPs (1 hr): Size Range 21 – 109 bp, and sequencing (2 hrs): 650 bp
Non Sanger-Based Sequencing Technology
The Illumina HiSeq 2000 is based on massively parallel sequencing of millions of nucleic acid fragments using reversible terminator-based sequencing chemistry. more. . .
Quantitative Real-Time PCR
The Applied Biosystems 7900HT Fast Real-Time PCR System is the only real-time quantitative PCR system that combines 96- and 384-well plate compatibility and the TaqMan® Low Density Array with fully automated robotic loading. The system also has the optional Fast Real-Time PCR capability. Currently, one ABI Prism 7900HT Sequence Detection System, three ABI Prism 7900HT Fast Real-Time PCR Systems and two StepOnePlus machines are available.
Digital Multiplexed Gene Expression Analysis Using the NanoString nCounter System
NanoString Technologies’ nCounter platform allows for highly multiplexed (20 to >550 targets per assay), digital quantification of nucleic acids. The platform requires no RT or amplification of template, both simplifying the workflow and eliminating potential bias, and requires only 100ng of RNA or a cell lysate from about 10,000 cells per assay. Since the system literally counts the individual nucleic acid molecules, it's perfect for downstream validation of RNAseq projects, as well as routine screening of pathways, array hits, or biomarker panels.