When writing dPCR materials and methods for a paper have you ever pondered what information you should include? This is where the MIQE guidelines will really help. Guidelines for minimum information required for publication of a digital PCR (dPCR) experiment were published by JF Huggett et al. in 2013. These were a follow-up to minimum information requirements for publication of real-time PCR experiments (qPCR) suggested by SA Bustin et al. in 2009. MIQE guidelines for qPCR and dPCR publications are divided into the following categories: essential information and desirable information. Essential Information must be included in the submitted manuscript or accompanying supplemental material. Desirable Information should be included in the submitted manuscript and is intended to help the reader understand the study.
MIQE guidelines are simple checklists intended to help researchers provide standardized information when publishing their quantitative PCR results. The goal is to help standardize nomenclature and decrease variability in the quality of published quantitative PCR data. MIQE got its name from the initial checklist published in 2009 for real-PCR experiments – Minimum Information for Publication of Quantitative real-time PCR Experiments (MIQE). A recently published update now provides a checklist for digital PCR – Minimum Information for Publication of Quantitative Digital PCR Experiments (dMIQE). MIQE checklist requirements specific to dPCR relate to reaction partitioning and data analysis.
What’s in the MIQE Guidelines?
MIQE guidelines include checklists of Essential Information and Desirable Information.
Essential Information must be submitted as part of your manuscript (body or supplemental materials).
Desirable Information should be submitted with your manuscript, if possible. This is information that would further help the reader understand your study.
Essential Information includes:
Experimental design, including controls
Sample description, microdissection or macrodissection, preparation & storage
Info on pseudogenes, retropseudogenes or other homologs if applicable
RTPrimerDB identification number
Probe sequence, if not proprietary
Oligo manufacturer and purification method
PCR buffer chemical constitution
Consumables catalog number & manufacturer
Data submission using RDML
Digital PCR-specific desirable information:
Gravimetric or volumetric dilutions (manual/robotic)
Total PCR reaction volume prepared
Partition volume variance/SD
Limit of detection of calibration control
Need an App for That?
Yes, there is an app for that! You may already be using the initial MIQE qPCR app released in 2011. An updated 2013 release now includes digital PCR checklists and digital PCR dedicated references (http://appshopper.com/reference/miqe-qpcr).
What’s the Benefit of Following MIQE Guidelines?
MIQE guidelines help standardize quantitative PCR experimental data, making it easier for other researchers to understand and replicate your study. That means more reliable data sharing and more breakthroughs for our scientific community. An example of a publication that has integrated them is Sedlak et al (2014) Identification of Chromosomally Integrated Human Herpesvirus 6 by Droplet Digital PCRClinical Chemistry v. 60, p.765-772.
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