Molecular Biology and Biochemistry Core

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Core C is the Molecular Biology and Biochemistry Core. This core is responsible for assisting each of the subprojects in microdissection of isolated kidney segments, genotyping, gene silencing, and in quantitation of mRNA and protein expression. Genotyping will be performed with real-time PCR using an ABI PRISM 7700 HT sequence detection system on mice in which the following genes have been disrupted: the p22phox subunit of NADPH oxidase; extracellular superoxide dismutase (EC-SOD), intracellular superoxide dismutase (IC-SOD), manganese superoxide dismutase (Mn- SOD), angiotensin type 1a receptor (AT-1aR), dopamine type 5 receptor (D5R), fibroblast growth factor type 2 (FGF-2) and type 7 (FGF-7) and the FGF binding protein (BP1) knockout and transgene. Core C will also assist in silencing the following genes by siRNA: BP1, FGF-2 and FGF- 7. In addition, Core C will assist in determining mRNA expression by real-time PCR, microarray and in situ hybridization and in assaying protein expression by Western blot, immunohistochemistry, radioligand binding and autoradiography in order to assess siRNA efficacy and specificity. This core will also be responsible for keeping investigators informed of the latest molecular techniques applicable for each of the subprojects.