Application Note
The MIQE Guidelines And Assessment Of Nucleic Acids Prior To qPCR And RT-qPCR
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•Application Note: The MIQE Guidelines And Assessment Of Nucleic Acids Prior To qPCR And RT-qPCR
By Thermo Scientific NanoDrop Products
Quantitative PCR has emerged as a fundamental technique in molecular & clinical diagnostics. Before now, little or no standardization in reporting existed, and as a result, experiments were frequently hard to replicate. In addition, many assumptions upon which the technique depends, in particular those pertaining to reference gene stability and amplification efficiency, were often in fact invalid. In order that experiments be better designed and appropriately validated, and that experimental reporting include sufficient information for experimental replication, the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines have been proposed (1). This application note examines the MIQE guidelines relevant to the assessment of nucleic acids, in particular quantification and purity assessment of nucleic acids using Thermo Scientific NanoDrop spectrophotometers.
The MIQE guidelines (1) are similar in principle to the MIAME guidelines used for microarray experiments. The rationale for the guidelines is that a full disclosure of methods allows assessment of protocol validity, correct interpretation of results and reproducibility. The guidelines cover all aspects of qPCR experiments, from experimental design to data analysis, although the bulk of the guidelines are associated with the experimental protocol itself.
Click Here To Download:•Application Note: The MIQE Guidelines And Assessment Of Nucleic Acids Prior To qPCR And RT-qPCR
