Evidence map›Paper›PMID 39407133›Full record

ArticleBMC bioinformatics2024

repDilPCR: a tool for automated analysis of qPCR assays by the dilution-replicate method.

Deyan Yordanov Yosifov, Michaela Reichenzeller, Stephan Stilgenbauer, Daniel Mertens

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Article in BMC bioinformatics, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

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4 · The record

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5 · Who and what money

Authors and funding

4 authors.

Deyan Yordanov YosifovDivision of CLL, Department of Internal Medicine III, Ulm University Hospital, Ulm, Germany. deyan.yosifov@uniklinik-ulm.de.
Michaela ReichenzellerDivision of CLL, Department of Internal Medicine III, Ulm University Hospital, Ulm, Germany.
Stephan StilgenbauerDivision of CLL, Department of Internal Medicine III, Ulm University Hospital, Ulm, Germany.
Daniel MertensDivision of CLL, Department of Internal Medicine III, Ulm University Hospital, Ulm, Germany.

Funding

Deutsche Forschungsgemeinschaft SFB1074, subprojects B1 and B2
6 · The paper itself

Abstract

backgroundThe dilution-replicate experimental design for qPCR assays is especially efficient. It is based on multiple linear regression of multiple 3-point standard curves that are derived from the experimental samples themselves and thus obviates the need for a separate standard curve produced by serial dilution of a standard. The method minimizes the total number of reactions and guarantees that Cq values are within the linear dynamic range of the dilution-replicate standard curves. However, the lack of specialized software has so far precluded the widespread use of the dilution-replicate approach.

resultsHere we present repDilPCR, the first tool that utilizes the dilution-replicate method and extends it by adding the possibility to use multiple reference genes. repDilPCR offers extensive statistical and graphical functions that can also be used with preprocessed data (relative expression values) obtained by usual assay designs and evaluation methods. repDilPCR has been designed with the philosophy to automate and speed up data analysis (typically less than a minute from Cq values to publication-ready plots), and features automatic selection and performance of appropriate statistical tests, at least in the case of one-factor experimental designs. Nevertheless, the program also allows users to export intermediate data and perform more sophisticated analyses with external statistical software, e.g. if two-way ANOVA is necessary.

conclusionsrepDilPCR is a user-friendly tool that can contribute to more efficient planning of qPCR experiments and their robust analysis. A public web server is freely accessible at https://repdilpcr.eu without registration. The program can also be used as an R script or as a locally installed Shiny app, which can be downloaded from https://github.com/deyanyosifov/repDilPCR where also the source code is available.

Indexed as

SoftwareReal-Time Polymerase Chain ReactionAutomationDilution-replicate designPlottingPolymerase chain reactionQPCRRRT-PCRShinyStatisticsWeb server

Identifiers

PMID39407133
PMCPMC11476982

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.