MIQE 2.0 Compliance: What You Need for Publication
What is MIQE 2.0?
The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines were first published in 2009 by Bustin et al. The 2025 update (MIQE 2.0) revises the Essential items list to reflect current best practices.
Many high-impact journals (Nature Methods, PLOS ONE, Journal of Molecular Diagnostics) require MIQE compliance or strongly recommend it. Submitting with a MIQE checklist signals rigor to reviewers.
Essential vs. Desirable items
MIQE items are categorized as Essential (must be reported for interpretable results) and Desirable (improve reproducibility). AnnealIQ focuses on the Essential items — these are what reviewers check.
Essential items cover: sample description, nucleic acid extraction, reverse transcription, qPCR target information, primers, protocol, validation, and data analysis.
What AnnealIQ auto-detects
AnnealIQ’s MIQE compliance card automatically populates items it can derive from your analysis state:
- • Reference genes used and validation method (after stability check)
- • Statistical tests applied
- • Normalization method (DDCt or Pfaffl)
- • Confidence intervals (when computed)
- • NTC/NRT control presence and results
- • Number of biological and technical replicates
- • Software and version used for analysis
These appear as green "Reported" items in the checklist.
Common gaps and how to fill them
The most common missing items are experimental details that only you know:
- • RNA extraction method: Tell the AI: "RNA was extracted using TRIzol reagent (Thermo Fisher) per the manufacturer’s protocol."
- • Reverse transcription details: "Reverse transcription used SuperScript IV (Thermo Fisher) with oligo(dT) primers, 1 μg total RNA input."
- • Primer sequences: "Forward: 5’-ATCGATCG-3’, Reverse: 5’-GCTAGCTA-3’, amplicon 120 bp."
- • Genomic DNA contamination assessment: This auto-populates if your data includes NRT controls.
Click the "Tell the AI" prompt chip next to any missing item to send the suggested message to the chat. The checklist updates automatically as you provide information.
Exporting your checklist
Click Copy on the MIQE card to get the full checklist as markdown, ready for a supplementary file. The checklist is also included in the FAIR data export bundle as miqe-checklist.md.
Tip: Fill in as many items as possible before exporting. A 90%+ complete checklist is a strong signal to reviewers that your qPCR data is reproducible.