The MIQE Guidelines
The MIQE guidelines define the minimum information necessary for proper evaluation of qPCR experiments and publications. By detailing all relevant experimental conditions and assay characteristics, the validity of the protocols used can be thoroughly assessed. Complete disclosure of all reagents, sequences, and analysis methods used enables other investigators to reproduce results.
The guidelines address several general categories including sample preparation, QC of nucleic acids, RT, qPCR assay, and data analysis. They also include an extensive checklist of “essential” or “desirable” items to be included in each of the following sections: experimental design, sample, nucleic acid extraction, RT, target information, primers and probe, qPCR protocol, qPCR validation and data analysis.
Most of the MIQE guidelines can be adopted quickly. Adoption of the “essential” items ensures that key variables affecting data quality are addressed, which, in turn, increases the degree of confidence in the results and the conclusions drawn. Adoption of some or all of the “desirable” items further ensures that a comprehensive approach is being taken to obtain quality data.
For instance, the guidelines recommend the disclosure of the probe sequence as highly desirable and strongly encouraged, but not all vendors of predesigned assays provide this information. Thermo Fisher Scientific provides probe sequences for all Solaris qPCR gene-expression assays.