160937 20251017144632.0 doi 10.1093/nar/gkaf386 sideral 144066 ART-2025-144066 eng Leonte, Radu Cristian IVA Prime: automated primer design for <i>in vivo</i> assembly cloning 2025 Access copy available to the general public Unrestricted Molecular cloning through in vivo assembly (IVA) is an efficient homology-based approach that can achieve complex cloning operations in a single step, bypassing the need for enzymes and reducing hands-on time and costs. However, primer design remains the most demanding task, requiring substantial expertise and a source of human error. Here, we introduce “IVA Prime,” an intuitive online tool for automated primer design for IVA cloning, which reduces the workload and avoids errors. We describe how IVA Prime generates optimized primers for all cloning operations, i.e. mutagenesis, insertions, deletions, and subcloning. IVA Prime provides an intuitive interface and robust algorithms, thus mitigating the complexities associated with primer design into a single mouse click. A set of thoroughly tested default parameters allows both inexperienced users and experts to generate ready-to-order primer sequences for a standardized cloning protocol with high success rate. We describe the method and demonstrate its efficiency and reliability with example applications. IVA Prime is freely available at https://www.ivaprime.com. info:eu-repo/grantAgreement/ES/MCINN/AEI/RYC2018-025720-I info:eu-repo/grantAgreement/ES/MICINN-AEI/PID2019-106284GA-I00 info:eu-repo/grantAgreement/ES/MICINN-FEDER/PID2020-113359GA-I00 info:eu-repo/grantAgreement/ES/MICINN/RYC2018-025731-I info:eu-repo/semantics/openAccess by-nc https://creativecommons.org/licenses/by-nc/4.0/deed.es info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Fischer, Patrick David Herguedas, Beatriz Universidad de Zaragoza (orcid)0000-0003-2044-4795 García-Nafría, Javier Universidad de Zaragoza (orcid)0000-0002-4254-3148 Hiller, Sebastian 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. (2025), [8 pp.] Nucleic acids res. Nucleic Acids Research 0305-1048 1516194 http://zaguan.unizar.es/record/160937/files/texto_completo.pdf Versión publicada 2314435 http://zaguan.unizar.es/record/160937/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:160937 articulos driver 2025-10-17-14:26:47 ARTICLE