Nucleic Acids

Instrumentation: Real-time qPCR instruments- Applied Biosystems 7900 (384-well) and Roche LC480 (96 or 384-well). We also have an ABI 7700s available to outside users with a flat fee/plate charge. Signup sheets are on the instruments.

RNA quality must be monitored prior to running real-time qPCR plate screening arrays (Exiqon, Roche, ABI, Qiagen, Lonza, etc). Samples are run on an Agilent 2100 biochip for quality control and must pass QC to be used for the succeeding real-time qPCR plate arrays. A Nanodrop instrument is available for outside users for an annual fee. We also have a Tecan Infinite 200 multimode plate reading spectrofluorometer multi-mode reader for fluorescent determination of DNA or RNA concentrations. Again, an annual user fee applies for this instrument for unlimted use. Prior to measuring miRNAs in serum/plasma quality is determined using a small number of miRNA assays. All real time qPCR assays are setup using roboitic liquid handlers, Tecan Genesis 100 (RT setup or additions of DNA samples) a Becman Biomek3000 (PCR master mix).

Assay modalities (all 384-well plate determinations except where mentioned below):

1. Nucleic acid detection and quantification, over 700 validated real-time qPCR assays available
   
2. miRNA screening ;Exiqon LNA-based primers, 2 384-well plate arrays for human, mouse and rat utilzing sensitive qPCR detection and GenEx software for data analysis. Focused arrays and custom arrays are also available.
   
3. Validation of miRNA (Exiqon), microarray, siRNA knockdown or compound screening‘hits’
   
4. Zygosity testing of mouse litters for endogenous or transgenes
   
5. Plate screening arrays from SA Biosciences/Qiagen, Lonza StellARrays, Life Technologies and Roche plate arrays for focused pathway or cell physiology determinations (96- or 384-well plates)
   
6. High resolution melt (HRM) analysis of SNPs, mutations, species differentiation, methylation and more. Inquire concerning this new application for real-time PCR.
   
7. Protein melt analysis for small compound/purified protein association screening studies. Can replace a fluorescence polarization assay if the association is strong enough, for less cost per assay.

Proteins

Instrumentation: MSD 2400 CCD camera electrochemoluminescence plate reader for MSD quantitative sandwich single or multiplex ELISA assay plates.

Biomek NXp 96-tip head liquid handling robotic workstation with integrated ThermoFisher Cytomat tip and plate store and Bio-tek ELx405 automated plate washer as well as a Biomek3000 for sample dispensing and setting up standard curves.

Assays: Any Meso Scale Discovery assay kit. Contact Dr. Shipley via phone at (713) 500-7458 or via e-mail at Gregory.L.Shipley@uth.tmc.edu for the latest information on kits. We can help with the development of novel assays on the MSD platform using single spot plates and reagents from MSD.

For information on Mass Spectrometry analysis of proteins and insight into protein purification and handling, contact Dr. Bill Dubinsky, Co-Director of the Proteomics Core Laboratory via e-mail at William.P.Dubinsky@uth.tmc.edu.