for the broader form of the guidelines. Other researchers have been creating further versions for specific forms of qPCR that may require a supplementary or different set of items to check, including single-cell qPCR and digital PCR (dPCR). Appropriate adherence to the existing MIQE guidelines has also been overviewed in other scientific areas, including photobiomodulation and clinical biomarkers. It was noted by Bustin in 2014 (and again by him in 2017) that there was some amount of uptake and usage of the MIQE guidelines within the scientific community, but there were still far too many published papers with qPCR experiments that lacked even the most basic of data presentation and proper confirmation of effectiveness for said data. These studies retained major reproducibility issues, where the conclusions of their evidence could not be replicated by other researchers, throwing the initial results into doubt. All of this was despite many papers directly citing Bustin's original MIQE publication, but not following through on the guideline checklist of material in their own experiments. However, some researchers have pointed out at least some success, with a number of papers being rejected by academic journals for publication due to failing to pass MIQE checklists. Other studies have been retracted after the fact once their lack of proper data to pass the MIQE guidelines was noted and publicly pointed out to the journal editors. Tightening of guidelines When setting up their new comparative qPCR systems titled "Dots in Boxes" in 2017, New England Biolabs stated that they had designed the data collection portion around the MIQE guidelines so that the data fit all the minimum parameter checklists in the protocols. Other scientific instrument companies have assisted in guideline compliance by purposefully tailoring their devices for them, including Bio-Rad creating a mobile app that allows for active marking of the MIQE checklist as each step is completed. An overview of the 10th anniversary since the publication of the MIQE guidelines was conducted in June 2020 and discussed the scientific studies that had produced better and more organized results when following the guidelines. In August 2020, an updated version of the guidelines for the digital PCR method was published to account for improvement in machinery, technologies, and techniques since the original 2013 release. Additional guideline steps were added for data analysis, while also providing a more simplified checklist table for researchers to use. An RT-qPCR targeting assay was developed