The use of real-time q PCR has nearly supplanted other approaches (e.g., Northern blotting, RNase protection assays). This review examines the current state of q PCR for gene expression analysis now that the method has reached a mature stage of development and implementation. A new quantitative PCR multiplex assay for rapid analysis of chromosome 17p11.2-12 duplications and deletions leading to HMSN/HNPP. The increasing number of patients with breast cancer whose survival has been improved by trastuzumab therapy, in the metastatic setting [1, 2] and probably also in the adjuvant setting [3, 4] underlines the need for highly reproducible yet cost-efficient methods for evaluating HER2.Selection of patients with HER2-overexpressing tumors for trastuzumab treatment is usually done on the basis of immunohistochemistry (IHC) combined with FISH for validation, which are at present the only the Food and Drug Administration-approved methods of HER2 analysis [IHC using the Hercept test (polyclonal antibody) or the m Ab CB11] .The Jo VE video player is compatible with HTML5 and Adobe Flash.Older browsers that do not support HTML5 and the H.264 video codec will still use a Flash-based video player. Bergqvist, Department of Oncology and Pathology, Radiumhemmet, Karolinska Institute and University Hospital, SE-171 76 Stockholm, Sweden.
Conclusion: The use of RNA-EP and Q-PCR to analyze HER2 in frozen and formalin-fixed breast cancer samples may be an alternate approach to IHC in combination with FISH/CISH.Specifically, the different fluorescent reporter technologies of real-time q PCR are discussed as well as the selection of endogenous controls. The conceptual framework for data analysis methods is also presented to demystify these analysis techniques. Since the founding of Bio Techniques, this journal has been a resource for the improvements in q PCR technology, experimental design, and data analysis. q PCR and, more specifically, real-time q PCR has become a routine and robust approach for measuring the expression of genes of interest, validating microarray experiments, and monitoring biomarkers. Date Published: 7/30/2011, Issue 53; doi: 10.3791/2777 Keywords: Molecular Biology, Issue 53, microarray, C.