Rheumatic Disease Research Core Center
The Rheumatic Disease Research Core Center is funded by the NIH through the University of Tennessee Health Science Center to provide core facilities that ensure cost effective, high quality, molecular and technical support for connective tissue research.
The Center of Excellence for Diseases of Connective Tissues maintains a core facility for the large-scale purification of collagen, and the production of alpha chains and CB peptides for research in autoimmunity. Tissues used as a source of cartilage are obtained exclusively from meat packing industry by-products.
Presently, these collagens are available from our core service at prices competitive with commercial entities providing this grade of product.
If you are interested in further information regarding pricing and availability
of collagen from our core service, please contact David Brand.
The mission at the UTHSC Animal Resource Core is to facilitate the study of rheumatic diseases by providing both expertise in mouse genetics and models of autoimmune arthritis to members of the Rheumatic Diseases Research Core Center (RDRCC) and other interested investigators.
Mouse Services
RDRCC Services:
- We perform backcrosses and set up mouse breeding programs.
- We screen using PCR or Flowcytometry for genotyping and to establish homozygosity.
- We offer the following mouse strains:
- DBA/1 Lac J Tg neg littermates (this stock is limited)
- DBA/1 Lac J Vb8.3
- DBA/1 Lac J a/b TCR Tg #24
- DBA/1 Lac J a/b TCR Tg #27
- B10.M-DR1 (DRB1*0101)
- B10.M-DR4 (DRB1*0401)
- DBA/1 IFN-g KO
- DBA/1LacJ a/bTCR Tg#24/IFN-gKO
- DBA/GFP- a/b TCR Tg #24
Flowcytometry Services
We have two flow cytometers available:
- FacsCalibur - a four color analyzer with a Multiwell Autosampler (MAS) for high throughput screening
- FacSort - a three color analyzer
We will perform or help you perform the multi-color immunofluorescent staining of cells for analysis.
Fluorochromes Available
Flowcytometry Screening Order Form
Bio-Plex
Multiplex Protein Analysis with a Bio-Plex/Luminex instrument
- Quantitates up to 100 proteins simultaneously from a single 100ul samples
- Analyzes Th1/Th2 cytokines, inflammatory mediators, chemokines,
and others - Murine or human samples
- Serum or tissue culture samples
- Kits available from a number of vendors
We will perform or help you perform the multi-color immunofluorescent staining of cells for analysis.
Vendors
BD Biosciences
BioRad
Biosource
Linco Research, Inc.
Luminex
To Schedule Use of the BioPlex
Call Laura or Nora at 901.523.8990 ext.7826
E-mail Laura at lmccorm1@uthsc.edu or Nora at nmoore@uthsc.edu.
Stop by the RDRCC/Animal Resource Core and sign up on our calendar posted outside
the door.
The Rheumatic Disease Research Core Center, funded by the NIH through UTHSC, is available to provide molecular and technical support for UTHSC investigators that are conducting research in rheumatic diseases. The lab provides a variety of services including: extracting and measuring RNA and DNA, providing quantitative real time Polymerase Chain Reaction (PCR) assays and analyzing single nucleotide polymorphism (SNP). The center can provide researchers with detailed information on genes and their expression.
Services
The RDRCC Molecular Resource Core Lab offers UTHSC researchers a wide range of genomic services starting from the extraction process, to developing primers and probes to analyzing polymerase chain reaction results. (Price List)
Extraction
RNA Extraction
The Core Lab provides the client with a variety of RNA extraction and purification protocols which include: Qiagen, Gentra Systems, Promega, and Trizol.
After extraction, the samples are measured for quality and quantity. We currently use the Gene Quant Pro Classic Spectrophotometer by GE Healthcare.
DNA Extraction
We are currently using DNeasy by Qiagen for DNA extraction and purification. This kit is designed for rapid isolation of total DNA from a variety of sample sources and is ideal for PCR.
Agilent 2100 Bioanalyzer
For a graphic representation of the quality of samples the Agilent 2100 is used. This is typically needed when the samples are being processed for cDNA microarrays which requires high quality RNA.
The 2100 bioanalyzer is a microfluidics based platform for measuring the quality of DNA, RNA, proteins and cells. It is the first commercial, analytical instrument based on lab-on-a-chip technology. One chip can run 12 samples. This provides an excellent alternative to messy, labor-intensive gels. The entire procedure takes two hours from start to finish. The data is presented in graph form.
Real Time PCR
The RDRCC Molecular Resource Core is equipped with the latest in PCR technology. We use the 7900HT Sequence Detection System developed by Applied Biosystems for detection and quantitation of PCR products.
Primers and probes can be designed, free of charge using Primer Express v2.0, for any gene whose nucleotide sequence is known. The RDRCC-MRC only charges for reagents, plates and the use of the 7900HT PCR instrument. See price list for more information.
We support Applied Biosystems Assays on Demand and Assays by Design for gene expression and SNP detection.
Low Density Array
The RDRCC-MRC now has the ability to run 7900HT TaqMan Low Density Array. The mirco fluidic card delivers TaqMan performance in a low density array format. The TaqMan chemistry delivers the greatest sensitivity, with over 6-logs of dynamic range and the highest specificity in quantitative gene expression. The user can select from over 40,000 inventoried TaqMan gene Expression Assays: gene-specific primer and probe sets for quantitative gene expression of human, mouse or rat genes.
Micro fluidic technology enables small, 2ul volume, 384-well reactions without the need for robotic pipetting instruments. TaqMan Arrays are delivered with your primer and probe already lyophilized in the wells.
Two microliter reactions dramatically reduce enzyme consumption. Micro fluidic technology prevents evaporation and eliminates small-volume pipetting errors.
| TaqMan Low Density Array Format Number | Number of Different Targets per Array + Control | Number of Assay Replicates | Number of Samples per Card | Part Number |
|---|---|---|---|---|
| 12 | 11+1 | 4 | 8 | 4342247 |
| 16 | 15+1 | 3 | 8 | 4346798 |
| 24 | 23+1 | 2(4) | 8(4) | 4342249 |
| 32 | 31+1 | 3 | 4 | 4346799 |
| 48 | 47+1 | 2(4) | 4(2) | 4342235 |
| 64 | 63+1 | 3 | 2 | 4346800 |
| 96a | 95+1 | 2(4) | 2(1) | 4342569 |
| 96b | 95+1 | 2(4) | 2(1) | 4342261 |
| 384 | 380+4 | 1 | 1 | 4342265 |
Freezer Plan
Our Applied Biosystem (ABI) freezer plan allows us to order reagents at a discount and pass the savings on to you. Some of the items included are: AmpliTaq, Sybr Green master mix, Random Hexamers, dNTP's, TaqMan PCR kits, and MicroAmp tubes. Call us for a complete list. You pay no shipping costs and discounts range from 13 to 30%. We will stock frequently ordered items.