Quantitative polymerase chain reaction (or qPCR) is a well-established assay for nucleic acid quantification and is still regarded as the method of choice in most areas of molecular biology. Though different types of qPCR quantification exist (absolute and relative), determining the amplification efficiency should be among the first things to do when setting up a qPCR assay. Understanding efficiency and how to calculate it is crucial for accurate data interpretation.
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Quantitative PCR (or qPCR), microarrays and RNA sequencing are all very valuable assays for in-depth gene expression analysis. But which one should you choose over the other for your next big experiment? Well quite frankly, it mostly depends on the goals of the project, your budget, and the organism of interest.
For many of us the process of pipetting transitioned into a subconscious routine to which we devote little thought after only a little practice, despite quality assurance placing great emphasis on liquid handling, pipette accuracy, repair and maintenance. Nevertheless, ensuring the competence of pipette operators (lab employees) is a way-too-often often neglected activity.
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The method of choice for nucleic acid (DNA, RNA) quantification in all areas of molecular biology is real-time PCR or quantitative PCR (qPCR). The method´s name derives from the fact that the amplification of DNA by polymerase chain reaction (PCR) is monitored in real time. It is a quantitative method in contrast to conventional PCR, meaning that it enables the determination of exact amounts (relative or absolute) of amplified DNA in samples.