Genomics
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Genomics covers a broad range of molecular techniques spanning the study of structure, function, evolution, mapping, and editing of genomes. Your core offers a number of technologies and services to meet your experimental needs.
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Genotyping
From snip to reportSimply provide us with your sample (2-3 mm tail snips or ear punches), primers, and a description of your validated PCR assay conditions. The Core Facility will then perform all the steps of the genotyping assay and send your report via email. The normal turn-around time for this service is two business days. The cost for the full genotyping service is $9/sample, competitive with outside vendors, PLUS there are no FedEx shipping fees.
DNA extraction only: For investigators who wish to do their own PCR reactions, but do not wish to expend valuable technician time and reagent costs for routine isolation of DNA, the Core Facility offers genomic DNA isolation services for $3/sample. the normal turn-around period for this service is one business day.
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Next-Generation Sequencing
Your core has partnered with the UT Genomics Core at the University of Tennessee Knoxville for access to an Illumina MiSeq high-throughput Next-Gen sequencer. The MiSeq platform provides up to 15 Gb of DNA sequence in a single run. Applications include genome sequencing, RNA sequencing, metagenomics, and 16S metabarcoding, as well as custom amplicon sequencing.
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qPCR
Real-time PCR, also called qPCR, RT-PCR, or kinetic PCR, is widely used in gene expression analysis either as a primary method or as a confirmation of other methods such as microarray analysis. Real-time PCR is also the most commonly used method to monitor the efficiency of gene silencing by RNA interference. Other applications include measurement of DNA copy number and transgene copy number, determination of viral titers, and single nucleotide polymorphism genotyping.
Unlike conventional PCR that uses an end-point analysis of the amplicon, real-time PCR is based on the detection and quantitation of the PCR product as it is made, using either fluorogenic specific probes (such as Taqman probes) or double-stranded DNA binding fluorescent dyes such as SYBR Green.
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Primer Design
Your core personnel can assist with primer design for PCR, qPCR, or sequencing. Contact us with details about your primer needs.
Publications
Phillips Campbell, R.B., Duffourc, M.M., Schoborg, R.V., Xu, Y., Liu, X., KenKnight, B.H., Beaumont, E. “Aberrant fecal flora
observed in guinea pigs with pressure overload is mitigated in animals receiving vagus
nerve stimulation therapy.” American Journal of Physiology- Gastrointestinal and Liver
Physiology. Oct 16, 311(4): G754-62. PMID: 27562060.
Slade, J.A, Hall, J.V., Kintner, J., Phillips-Campbell, R., Schoborg, R.V. “Host nectin-1 promotes chlamydial infection in the female mouse genital tract, but is not required for infection in a novel male murine rectal infection model.” PLOS One. Aug 16, 11(8): e0160511. PMID: 27486990.
Ordway GA, Szebeni A, Chandley MJ, Stockmeier CA, Xiang L, Newton SS, Turecki G, Duffourc M, Zhu MY, Zhu H, and Szebeni K. (2012). Low gene expression of bone morphogenetic protein 7 in brainstem astrocytes in major depression. Int J of Neuropsychopharmacol 15(7):855-68; epub 2011 Sep6:1-14. PMID: 21896235
Fan Y, Huang J, Duffourc M, Kao R, Ordway G, Huang R, and Zhu M-Y. (2011). Transcription factor Phox2 upregulates expression of norepinephrine transporter and dopamine β-hydroxylase in adult rat brains. Neuroscience 192:37-53. Epub 2011 Jul 16. PMID: 21763404.
Brown RW, Noel DM, Smith JJ, Smith ML, Huggins KN, Szebeni K, Szebeni A, Duffourc M, Chandley M, and Ordway GA. (2011) Eszopiclone facilitation of the antidepressant efficacy of fluoxetine using a social defeat stress model. Pharmacol Biochem Behav epub 2011 Jun 15. PMID: 21699914
Campbell SE, Rudder B, Phillips RB, Whaley SG, Stimmel JB, Leesnitzer LM, Lightner J, Dessus-Babus S, Duffourc M, Stone WL, Newman RA, Yang P, Aggarwal BB, and Krishnan K. (2011) Gamma tocotrienol induces growth arrest through a novel pathway involving TGFb2 in prostate cancer. Free Radic Biol Med 50(10):1344-54. Epub 2011 Feb 16. PMID: 21335085
Stuart CA, Howell ME, Baker JD, Dykes RJ, Duffourc MM, Ramsey MW, Stone MH. (2010) Cycle Training Increased GLUT4 and Activation of mTOR in Fast Twitch Muscle Fibers. Med Sci Sports Exerc 42(1):96-106.
Campbell SE, Musich PR, Whaley SG, Stimmel JB, Leesnitzer LM, Dessus-Babus S, Duffourc M, Stone W, Newman RA, Yang P, Krishnan K. (2009) Gamma Tocopherol Up regulates the Expression of 15-S-HETE and Induces Growth Arrest Through a PPAR γ-Dependent Mechanism in PC-3 Human Prostate Cancer Cells. Nutr Cancer 61(5):649-62.
Ordway, G.A., Szebeni, A., Duffourc, M.M., Dessus-Babus, S. and Szebeni, K. (2009) Gene Expression Analyses of Neurons, Astrocytes and Oligodendrocytes Isolated by Laser Capture Microdissection from Human Brain: Detrimental Effects of Laboratory Humidity. J. Neuroscience Research. 87(11):2430-8.
Vanover, J., Sun, J., Deka, S., Kintner, J., Duffourc, M. and R.V. Schoborg. (2008) Herpes Simplex Virus Co-infection Induced Chlamydia trachomatis Persistence is not Mediated by any Known Persistence Inducer or Anti-chlamydial Pathway. Microbiology 154:971-978.
Mabe, Abigail M., Hoard, Jennifer L., Duffourc, Michelle M., and Hoover, Donald B. (2006) Localization of cholinergic innervation and neurturin receptors in adult mouse heart and expression of the neurturin gene. Cell & Tissue Research 326(1):57-67