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PCRdrive: the largest qPCR assay archive to date and endless potential for lab workflow revitalization

BACKGROUND: Primer design is a crucial step in establishing specific and sensitive qPCR assays. Even though numerous tools for primer design exist, the majority of resulting assays still requires extensive testing and optimisation or does not allow for high quality target amplification. We developed...

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Autores principales: Burger, Florian, Angioni, Michele, Russo, Gianluca, Schad, Martina, Kallarackal, Jim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6249872/
https://www.ncbi.nlm.nih.gov/pubmed/30466398
http://dx.doi.org/10.1186/s12859-018-2452-3
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author Burger, Florian
Angioni, Michele
Russo, Gianluca
Schad, Martina
Kallarackal, Jim
author_facet Burger, Florian
Angioni, Michele
Russo, Gianluca
Schad, Martina
Kallarackal, Jim
author_sort Burger, Florian
collection PubMed
description BACKGROUND: Primer design is a crucial step in establishing specific and sensitive qPCR assays. Even though numerous tools for primer design exist, the majority of resulting assays still requires extensive testing and optimisation or does not allow for high quality target amplification. We developed a workflow for designing qPCR assays. Unlike other tools, we compute a PCR assay including primer design, concentrations and the optimal PCR program. RESULTS: Gene expression assays were already generated in a total of 283,226 genes from three species and are continued for all genes of the major model species. The results are available online at https://pcrdrive.com/lab#/assay-database. The workflow involves filtering Primer3-generated primers by considering diverse parameters including specificity, single-nucleotide polymorphisms (SNPs), secondary structure as well as compatibility with standard qPCR assay conditions. The resulting assays consist of transcript-specific primer sequences, a reagents protocol as well as instrument settings which are provided in a web-based tool called PCRdrive. PCRdrive was designed to support PCR users in their PCR-related tasks and is equipped with handy functions, components of an electronic lab notebook (ELN) as well as teamworking opportunities. CONCLUSION: High quality ready to use qPCR assays for gene expression analysis are provided within the online platform PCRdrive. A built-in primer designer enables easy generation of assays which is not supported by any other tool. The wet lab optimisation of new assays can be transparently documented and shared within the team. PCRdrive also contains an archive of public PCRs which is updated regularly. Users may use the archive to publish their PCR to the community which makes it easy for other researchers worldwide to reproduce and validate the PCR. PCRdrive is a growing network of PCR users, simplifying and streamlining research through its useful existing features and continuous developments from the active development team.
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spelling pubmed-62498722018-11-26 PCRdrive: the largest qPCR assay archive to date and endless potential for lab workflow revitalization Burger, Florian Angioni, Michele Russo, Gianluca Schad, Martina Kallarackal, Jim BMC Bioinformatics Software BACKGROUND: Primer design is a crucial step in establishing specific and sensitive qPCR assays. Even though numerous tools for primer design exist, the majority of resulting assays still requires extensive testing and optimisation or does not allow for high quality target amplification. We developed a workflow for designing qPCR assays. Unlike other tools, we compute a PCR assay including primer design, concentrations and the optimal PCR program. RESULTS: Gene expression assays were already generated in a total of 283,226 genes from three species and are continued for all genes of the major model species. The results are available online at https://pcrdrive.com/lab#/assay-database. The workflow involves filtering Primer3-generated primers by considering diverse parameters including specificity, single-nucleotide polymorphisms (SNPs), secondary structure as well as compatibility with standard qPCR assay conditions. The resulting assays consist of transcript-specific primer sequences, a reagents protocol as well as instrument settings which are provided in a web-based tool called PCRdrive. PCRdrive was designed to support PCR users in their PCR-related tasks and is equipped with handy functions, components of an electronic lab notebook (ELN) as well as teamworking opportunities. CONCLUSION: High quality ready to use qPCR assays for gene expression analysis are provided within the online platform PCRdrive. A built-in primer designer enables easy generation of assays which is not supported by any other tool. The wet lab optimisation of new assays can be transparently documented and shared within the team. PCRdrive also contains an archive of public PCRs which is updated regularly. Users may use the archive to publish their PCR to the community which makes it easy for other researchers worldwide to reproduce and validate the PCR. PCRdrive is a growing network of PCR users, simplifying and streamlining research through its useful existing features and continuous developments from the active development team. BioMed Central 2018-11-22 /pmc/articles/PMC6249872/ /pubmed/30466398 http://dx.doi.org/10.1186/s12859-018-2452-3 Text en © The Author(s) 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Software
Burger, Florian
Angioni, Michele
Russo, Gianluca
Schad, Martina
Kallarackal, Jim
PCRdrive: the largest qPCR assay archive to date and endless potential for lab workflow revitalization
title PCRdrive: the largest qPCR assay archive to date and endless potential for lab workflow revitalization
title_full PCRdrive: the largest qPCR assay archive to date and endless potential for lab workflow revitalization
title_fullStr PCRdrive: the largest qPCR assay archive to date and endless potential for lab workflow revitalization
title_full_unstemmed PCRdrive: the largest qPCR assay archive to date and endless potential for lab workflow revitalization
title_short PCRdrive: the largest qPCR assay archive to date and endless potential for lab workflow revitalization
title_sort pcrdrive: the largest qpcr assay archive to date and endless potential for lab workflow revitalization
topic Software
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6249872/
https://www.ncbi.nlm.nih.gov/pubmed/30466398
http://dx.doi.org/10.1186/s12859-018-2452-3
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