Revolutionizing Molecular cloning: Introducing FastCloneAssist, a Streamlined Python tool for optimizing primer design in restriction & ligation-independent PCR cloning.
FastCloning, a paradigm shift in PCR cloning, has streamlined the process by eliminating laborious, multi-step traditional methods. This innovative technique, pioneered by Li et al. (2011), utilizes overlapping PCR primers and DpnI digestion for seamless integration of insert DNA into any desired ve...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2025-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0306950 |
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| author | Pradip Kumar Singh Michael S Donnenberg |
| author_facet | Pradip Kumar Singh Michael S Donnenberg |
| author_sort | Pradip Kumar Singh |
| collection | DOAJ |
| description | FastCloning, a paradigm shift in PCR cloning, has streamlined the process by eliminating laborious, multi-step traditional methods. This innovative technique, pioneered by Li et al. (2011), utilizes overlapping PCR primers and DpnI digestion for seamless integration of insert DNA into any desired vector position, regardless of restriction sites. This versatility makes FastCloning ideal for constructing fusion proteins, chimeric cDNAs, and manipulating genes with unparalleled ease. However, efficient primer design remains a critical hurdle, particularly for newcomers, as errors can lead to failed cloning attempts. To address this bottleneck, we present FastCloneAssist, a user-friendly Python program that automates FastCloning primer design with minimal user input. Users simply provide vector and insert sequences, along with the desired melting temperature (Tm), and FastCloneAssist provides best primer pairs after calculating optimal primer parameters for efficient PCR amplification and seamless DNA integration using established bioinformatics libraries. This open-source, freely available tool simplifies and accelerates cloning, making this powerful technique accessible to researchers of all levels and expediting scientific discovery. |
| format | Article |
| id | doaj-art-2f21bdfada0441fa8aff8ef490bbb24e |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-2f21bdfada0441fa8aff8ef490bbb24e2025-08-20T01:54:21ZengPublic Library of Science (PLoS)PLoS ONE1932-62032025-01-01203e030695010.1371/journal.pone.0306950Revolutionizing Molecular cloning: Introducing FastCloneAssist, a Streamlined Python tool for optimizing primer design in restriction & ligation-independent PCR cloning.Pradip Kumar SinghMichael S DonnenbergFastCloning, a paradigm shift in PCR cloning, has streamlined the process by eliminating laborious, multi-step traditional methods. This innovative technique, pioneered by Li et al. (2011), utilizes overlapping PCR primers and DpnI digestion for seamless integration of insert DNA into any desired vector position, regardless of restriction sites. This versatility makes FastCloning ideal for constructing fusion proteins, chimeric cDNAs, and manipulating genes with unparalleled ease. However, efficient primer design remains a critical hurdle, particularly for newcomers, as errors can lead to failed cloning attempts. To address this bottleneck, we present FastCloneAssist, a user-friendly Python program that automates FastCloning primer design with minimal user input. Users simply provide vector and insert sequences, along with the desired melting temperature (Tm), and FastCloneAssist provides best primer pairs after calculating optimal primer parameters for efficient PCR amplification and seamless DNA integration using established bioinformatics libraries. This open-source, freely available tool simplifies and accelerates cloning, making this powerful technique accessible to researchers of all levels and expediting scientific discovery.https://doi.org/10.1371/journal.pone.0306950 |
| spellingShingle | Pradip Kumar Singh Michael S Donnenberg Revolutionizing Molecular cloning: Introducing FastCloneAssist, a Streamlined Python tool for optimizing primer design in restriction & ligation-independent PCR cloning. PLoS ONE |
| title | Revolutionizing Molecular cloning: Introducing FastCloneAssist, a Streamlined Python tool for optimizing primer design in restriction & ligation-independent PCR cloning. |
| title_full | Revolutionizing Molecular cloning: Introducing FastCloneAssist, a Streamlined Python tool for optimizing primer design in restriction & ligation-independent PCR cloning. |
| title_fullStr | Revolutionizing Molecular cloning: Introducing FastCloneAssist, a Streamlined Python tool for optimizing primer design in restriction & ligation-independent PCR cloning. |
| title_full_unstemmed | Revolutionizing Molecular cloning: Introducing FastCloneAssist, a Streamlined Python tool for optimizing primer design in restriction & ligation-independent PCR cloning. |
| title_short | Revolutionizing Molecular cloning: Introducing FastCloneAssist, a Streamlined Python tool for optimizing primer design in restriction & ligation-independent PCR cloning. |
| title_sort | revolutionizing molecular cloning introducing fastcloneassist a streamlined python tool for optimizing primer design in restriction amp ligation independent pcr cloning |
| url | https://doi.org/10.1371/journal.pone.0306950 |
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