The beginning of cell death due to the production of toxic modified SWS. In this case, additional SWS could ameliorate the first, but enhance the latter thereby not having an overall effect. However, the acute effect could also be due to TOCP acting on another target, an aspect that has to be explored in the future. Reverse transcription quantitative real-time polymerase chain reaction tests that measure transcript abundance of selected genes in clinical specimens have been demonstrated to increase cancer diagnostic accuracy and Gambogic-acid enable “personalized medicine” through selection of the most effective treatment for each cancer. However, a key challenge is that current clinical pathology sample collection and processing procedures focus on formalin fixation and paraffin embedding and fresh/fresh-frozen tissues rarely are available for molecular analysis. FFPE samples are difficult to work with because they yield RNA that 1) often contains PCR-interfering substances, and 2) is uniformly highly fragmented and often in low abundance. Economic factors prevent changing this workflow to ensure collection of samples in a form more conducive to molecular genetic analysis, such as fresh frozen, therefore, there is a need to develop methods that are sufficiently robust to reliably conduct molecular genetic analysis in FFPE samples. One way to address the challenge of interfering substances is to incorporate quality control in qPCR through measurement of each analyte relative to a known number of competitive template internal standard copies. This quality control method is recommended by regulatory agencies, including the EPA, ISO, and FDA, and is implemented in many Tubeimoside-I FDA-approved single analyte RT-qPCR tests. Multiple gene RT-qPCR tests present an even more complex quality control challenge that can be addressed by combining the IS for each of the genes into an internal standards mixture and using an aliquot of this ISM in each experiment, as previously described. Each target gene and loading control gene then is measured relative to a known number of its respective competitive template IS molecules in each PCR reaction. To address the challenge of analyzing clinical samples that yield a low amount of RNA, we previously described a competitive multiplex PCR method, in which all reference and target genes are first co-amplified with ISM in a first round of PCR, followed by amplification of individual gene in the second round. This approach enables reliable measurement of many genes from the amount of RNA that, without pre-amplification, would be sufficient for measurement of only a single gene. The primary goal of this study was to develop a robust RTqPCR method for more reliable molecular diagnostic testing of FFPE samples including those stored in existing large archives. To meet this need, a method was designed with four elements: 1) Synthetic competitive IS formulated into an ISM to control for sub-optimal PCR, including interference with PCR caused by substances present in FFPE samples, sub-optimal quantity or quality of PCR reaction reagents, and inter-well and/or interplatform variation in PCR conditions; 2) Fluorometric hydrolysis probe real-time PCR to enable quantification of short PCR amplicons that are optimal for reliable analysis of FFPE samples; 3) An external standards mixture in each experiment to control for inter-lot and inter-experimental variation in probe fluorescence intensity; and 4) Reverse transcription with gene specific primers followed by competitive multiplex pre-amplification.