Molecular Resource Center Equipment

Bio-Tek FLX 800TBE Microplate Fluorescence Reader
DNA Sequencing and Fragment Analysis
Fluorescence Microscopy
Illumina Microarray BeadStation
Universal Probe Library
Liquid Handling Robot
Microarrays
Phosphor-imaging

Bio-Tek FLX 800TBE Microplate Fluorescence Reader

This microplate reader can detect fluorescence and luminescence at both endpoint and kinetic readings in 6, 12, 24, 48, 96 and 384-well plates. The reader has an excitation range of 300-650nm and emission range of 340-800nm. The MRC currently has 360nm, 485nm and 508nm excitation filters as well as 460nm, and 528nm emission filters. Training can be provided for this equipment.

DNA Sequencing and Fragment Analysis

The MRC currently has an ABI PRISM 310 and two ABI PRISM 3130XL Genetic Analyzers for DNA sequencing and fragment analysis. The ABI 3130XL’s has 16 capillaries that operate in parallel and are fully automated from sample loading to data analysis. We currently employ two of these instruments; one devoted to sequencing and the other dedicated to fragment analysis. The 310 is a single capillary instrument that can support most gel-based assays. It is currently used for fragment analysis employing fluorescent markers that precede the ones used with the 3130XL. As part of our DNA sequencing services, we offer plasmid amplification (Templiphi) for sequencing only. Details on submission of transformed bacteria and costs can be obtained from the MRC.


Zeiss AxioPhot

Fluorescence Microscopy

The Axiophot Photomicroscope is available for either transmitted light or fluorescence detection on stained slides. Optronics Magnafire software can be used to capture and print images, using a digital camera, from bright field monochrome (grayscale) images and color images as well as fluorescent images.

Illumina Microarray BeadStation

The MRC in collaboration with the Vision Research Core of the Hamilton Eye Institute is pleased to make available an Illumina whole genome SNP and Microarray gene expression analysis core service at UTHSC.Gene expression analysis includes RNA QA/QC, RNA labeling, processing of chip and data analysis.The BeadStation is a complete solution featuring microarrays configured for whole genome gene expression, standard or custom focused arrays for gene expression. Convenient multi-sample array formats reduce array handling requirements and allow researchers to benefit from lower variability and increased experimental throughput. This equipment affords the researcher an entirely new level of quality, flexibility and throughput for gene expression profiling. It enables Direct Hybridization gene expression into a multi-use genetic analysis system that can also do GoldenGate Genotyping.Contact Dr. William Taylor at 448-6191 for additional information.


LightCycler^®480

Universal Probe Library

The LightCycler^® 480 Instrument is a rapid, thermal blockcycler with integrated real-time, online detection capabilities. Based on a comprehensive improvement of the Peltier-based block technology, the Instrument provides extraordinary well-to-well temperature homogeneity and maximized inter-well, inter-cycle reproducibility. The special arrangement of optical components in the LightCycler^® 480 Instrument ensures the uniform collection of signals across the plate and makes analysis independent of the sample position on the plate.The LightCycler^® 480 System setup enables the use of all current probe formats (e.g., SYBR Green I, Hydrolysis Probes, HybProbe Probes) and provides the ideal solution for fast and precise qualitative or quantitative detection of nucleic acids, genotyping, and mutation analysis. The ability to freely combine five excitation and six emission filters permits analysis of signals from multiple dyes (e.g., LightCycler^® Red 610, 640, LightCycler^® CYAN 500, FAM, VIC, HEX, Cy5, and Fluorescein) in monocolor as well as in multiplex assays.The user-friendly LightCycler^® 480 Software enables highly flexible and extensive data analysis for different scientific needs. Based on the modular design of the software, the user can customize the system by adding advanced analysis modules (e.g., LightCycler^® 480 Relative Quantification Software, LightCycler^® 480 Genotyping Software) to the LightCycler^® 480 Basic Software (run option, Tm calling option, absolute quantification option).

Liquid Handling Robot

The MRC has excess capacity for all of our services. A liquid handling robot (Perkin Elmer Multiprobe HT EX) can be used to scale up assays as necessary. We are currently capable of sequencing 300 samples per day and performing >3000 SNPs per day utilizing existing technology. Upgrading the equipment in the lab to 384 well versions would increase this number to >15,000 SNPs per day. Finally, with newer multiplex techniques (SNplex from Applied Biosystems), available soon, we can provide greater than 70,000 SNP genotypes per day. Microarrays can be produced for any species provided a DNA library is available. This is important for this proposal since bacterial species will be used for which no commercial microarrays are available.


Agilent Bioanalyzer       Gene Pix 4000B

Microarrays

The production of microarrays is currently offered as a service by the MRC. DNA for the arrays can be produced as either amplified cDNA clones or oligonucleotides and applied to glass substrates. Users must supply the appropriate cDNA clones or sequences for the oligonucleotides. Software is available to aid the user in the design of the appropriate oligonucleotides. Currently, a mouse 15K array from cDNA clones supplied by the National Institute of Aging is available, along with a specific array for a number of neural specific genes from rat. Bacterial genomic DNA arrays for Salmonella and Streptococcus are also available. As part of the microarray facility, an Agilent Bioanalyzer is also available in the MRC. The Bioanalyzer system is an automated, computer controlled, integrated analyzer capable of sizing and concentration determinations of nucleic acids and proteins. Sample preparations include PCR and RT-PCR fragments, restriction digests, total RNA and mRNA, cell lysates, column fractions and purified proteins. The primary use of the Bioanalyzer is to determine the quality of RNA for microarray analysis. Finally, facilities for data collection from microarrays also exist employing an Axon Instruments 4000B confocal microarray scanner with Gene Pix Software.


Molecular Dynamics Typhoon 941
(located at the VAMC – Room BB107)

Phosphor-imaging

The Molecular Dynamics Typhoon Phosphor-imager is capable of scanning exposed storage phosphor screens, fluorescent (532nm or 633nm excitation), and chemiluminescent samples. ImageQuant software analysis tools can be used to perform area or volume quantification on scanned images. DNA and protein fragments separated by electrophoresis can be analyzed using the fragment analysis software.


The QIAcube is designed to perform fully automated purification of nucleic acids and proteins for research use in molecular biology applications. Up to 12 samples can be processed per run using proven QIAGEN spin-column technology. The instrument controls integrated components including a centrifuge, heated shaker, pipetting system, and robotic gripper. The system is a hands-off system and already contains a centrifuge.


SpectraMax M2e Microplate/Cuvette Reader
The M2e Multi-Mode Spectrophotometer is used for fluorescence intensity (top and bottom reading), UV/Vis absorbance, luminescence (secondary mode), & TRF (secondary mode) reading modes, tunable wavelength selection for excitation & emission, built-in cuvette port and microplate drawer, precision endpoint/kinetic/spectral scanning/well scanning modes, proprietary PathCheck Sensor, thermal regulation from ambient + 4 oC-45 oC.

Contact Us

Don Thomason, Ph.D.
Molecular Resource Center
360 Cancer Research Building
Memphis, TN 38163

Phone: (901) 448-2656
Email: dthomaso@uthsc.edu