Today, quantitative reverse transcription polymerase chain reaction (qRT-PCR) is the favored method to quantify gene expression in molecular biology and clinical studies. qRTPCR analyses can be performed in a high-throughput manner and are very sensitive and specific, cost-effective, and reproducible. Even though this technique is easy to handle, the executer has to pay attention to some critical points. Right from the beginning of proper data generation, it is important to store the samples correctly and to prepare pure and intact RNA. It is, for example, more difficult to obtain good-quality RNA from paraffin-embedded samples than of frozen tissues (Takano et al. 2010). There are RNA protection and preparation kits available that help to maintain the integrity of RNA. Subsequently, an effective reverse transcription reaction (RT) and a specific primer design are indispensable for a high PCR efficiency. For a clear survey, Udvardi's group posted 11 golden rules of performing qRT-PCR analyses and gave a concise support from sample collection to data analysis (Udvardi et al. 2008). Before performing qRT-PCR analyses, the choice between two detection methods has to be made: nonprobe-based and probe-based assays. Nonprobe-based analyses use an intercalating dye like SYBR® green or ethidium bromide for the target detection. The dye intercalates into all double stranded DNAs formed during the PCR reaction. To exclude the detection of unspecific PCR products, it is necessary to analyze the PCR products further bymelting curves or gel electrophoresis. Themajor advantage of nonprobe-based analyses even in high throughput experiments is the lower investment, because each optimized PCR can easily be converted into a quantitative PCR. In contrast, probe-based detection needs an additional oligonucleotide that binds internal to the amplified target sequence and contains a quencher and reporter. The probe has to be designed for every target sequence, and this, in turn, guarantees a higher specificity for the detection system. Several different probe structures can be used for this method: TaqMan®, molecular beacons, or Scorpion Primers. A crucial point of qRT-PCR analyses is the normalization of the results that, improperly done, can have a profound influence on study conclusions (Ferguson et al. 2010). Even though normalization is a highly discussed topic, mostly neither editors nor reviewer demands on the proof of a suitable data evaluation. Current used normalization strategies range from standardization of sample size, like tissue weight or RNA amount, to internal and external standards. When normalizing with external standards, the absolute copy number of the target transcripts can be determined. The standard curve can either be generated from diluted plasmid DNA (Li and Wang 2000) or from in vitro-transcribed RNAs (Workenhe et al. 2008). The quantification is based on the assumption that plasmid DNA and cDNA [reverse transcribed frommessenger RNA (mRNA)] have the same amplifications rates. In contrast to the external control, it is also possible to use coamplified internal controls. This normalization strategy to one or more internal controls is commonly used (Vandesompele et al. 2002). The principle of this method is based on the quantification of the mRNA of an internal reference gene that undergoes the same procedures and conditions as the mRNA of interest. Using this kind of normalization, it is absolutely essential to search for a reference gene whose expression is Franziska Bollmann, Ingrid Casper, and Jenny Henke contributed equally to this editorial.