References:
- Ran Xie, David S. Loose, Gregory L. Shipley, Susu Xie, Roland L. Bassette Jr., and Russell R. Broaddus (2007). Hypomethylation Induced Expression of S100A4 in Endometrial Carcinoma, Modern Pathology, 20, 1045-1054.
- Shipley, G.L. (2006) Chapter 1: An Introduction to Real-Time PCR. In Real-Time PCR (Dorak MT (Ed), New York, Taylor & Francis Group.
- Restrepo, B.I., D.I. Gomez, G.L. Shipley, J.B. McCormick and S.P Fisher-Hoch (2006). Selective enrichment and detection of mycobacterial DNA in paucibacillary specimens, J. Microbiol. Methods, 67, 220-229
- Wiley-VCH Verlag, Weinheim. Deng, L., R.R. Broaddus, A. McCampbell, G.L. Shipley, D.S. Loose, G.M. Stancel, J.H. Pickar and P.J.A. Davies (2005). Identification of a Novel Estrogen-Regulated Gene, EIG121, Induced by Hormone Replacement Therapy and Differentially Expressed in Type I and Type II Endometrial Cancer, Clin Cancer Res, 11, 8258-8264.
- Shipley, G.L. (2005) Real-Time Quantitative PCR: Theory and Practice. In Encyclopedia of molecular cell biology and molecular medicine (R. A. Meyers, Ed.), Vol. 11, pp. 481-523.
- K.K. Sra, M. Babb-Tarbox, S. Aboutalebi, P. Rady, G.L. Shipley, D.D. Dao and S.K. Tyring (2005). Molecular Diagnosis of Cutaneous Diseases, Arch. Dermatol., 141, 225-241.
- Williams, Malgorzata, L., S.A. Wrighton, P.J.A. Davies, D.S. Loose, G.L. Shipley and H.W. Strobel (2004). Estrogen Regulation of the Cytochrome P450 3A Subfamily in Humans. J. Pharm. Exp. Therapeutics, 311(2), 728-735.
- Yeung, A.T., Holloway, B.P., Adams, P.S. and Shipley, G.L. (2004). Evaluation of dual-labeled fluorescent DNA probe purity versus performance in real-time PCR, Biotechniques, 36, 266-275.E.T
Greg L. Shipley , Ph.D.
Assistant Professor
UTHSC, Medical School, (713) 500 - 7458
Gregory.L.Shipley@uth.tmc.edu
Quantitative Genomics Core Laboratory
http://research.uth.tmc.edu/corelabs/qgcl.html
The Quantitative Genomics Core Laboratory provides high throughput processing and analysis for two distinct technologies; 1- real-time quantitative PCR and 2- quantitative ELISA determination. Utilizing real-time qPCR or RT-qPCR, nucleic acids purified from cellular, viral or microbial sources can be detected and quantified. We also have employed methods to perform real-time RT-qPCR from whole cell extracts without RNA purification. The laboratory is setup for high-throughput sample analysis through the use of Tecan and Beckman liquid handling robotics. Real-time qPCR analysis is performed using Applied Biosystems 7700 and 7900 sequence detectors and the Roche LC480 platforms. Both 96- and 384-well plates can be accommodated depending on the number of samples to be analyzed. Each plate contains a 5-log standard curve and a no template control. RNA samples are run in triplicate with reverse transcriptase plus a fourth well without reverse transcriptase as a DNA contamination control. DNA samples are run in triplicate or, for zygosity testing, as 6 replicates. Up to 21 RNA or 28 DNA samples are run per 96-well plate while up to 93 samples can be run per 384-well plate. We can also have plate configurations for two assays side-by-side on one 384-well plate with up to 45 samples/assay. If the over 500 existing probe-based assays do not include your transcript or gene of interest, we can design and QC new real-time qPCR assays (Taqman probe-based assays) for you as well as those utilizing the new Roche Universal Probe Library. We can also perform miRNA analyses on RNA samples utilizing kits from Applied Biosystems. We have over 11 years experience in designing assays and performing real-time qPCR analysis on nucleic acids. Other applications of the technology are microarray validation and the quantification of miRNAs. Protein quantification is accomplished utilizing a MesoScale electrochemiluminescent plate reader. The plates have a capture antibody for binding the target antigen and a second antibody is used for detection. Up to 10 different assays can be multiplexed in a single well of a 96-well plate. Plates, samples and reagents are manipulated using a liquid handling robot (Beckman NXp) and an automated plate washer (Biotech ELx405) to ensure uniform handling. Many assays are available from MesoScale and more are being developed continuously.
