{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Zhao M"],"funding":["Northwest Center for Small Fruits Research","Ministerio de Ciencia e Innovación","Inner Mongolia Agricultural University","NHGRI NIH HHS","National Natural Science Foundation of China","National Institutes of Health"],"pagination":["36"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC7990137"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["2"],"pubmed_abstract":["An unprecedented number of viruses have been discovered by leveraging advances in high-throughput sequencing. Infectious clone technology is a universal approach that facilitates the study of biology and role in disease of viruses. In recent years homology-based cloning methods such as Gibson assembly have been used to generate virus infectious clones. We detail herein the preparation of home-made cloning materials for Gibson assembly. The home-made materials were used in one-step generation of the infectious cDNA clone of a plant RNA virus into a T-DNA binary vector. The clone was verified by a single Illumina reaction and a de novo read assembly approach that required no primer walking, custom primers or reference sequences. Clone infectivity was finally confirmed by <i>Agrobacterium</i>"],"journal":["Phytopathology research"],"pubmed_title":["Home-made enzymatic premix and Illumina sequencing allow for one-step Gibson assembly and verification of virus infectious clones."],"pmcid":["PMC7990137"],"funding_grant_id":["U24 HG006620","BIO2016-80572-R","HG006620","U41 HG006620","NDGCC2016-23","31860489","PID2019-109380RBI00 / AEI / 10.13039/501100011033 (AEI-FEDER)"],"pubmed_authors":["Zhao M","Tzanetakis IE","Simon-Mateo C","Gallo A","Garcia JA","Pasin F","Garcia B"],"additional_accession":[]},"is_claimable":false,"name":"Home-made enzymatic premix and Illumina sequencing allow for one-step Gibson assembly and verification of virus infectious clones.","description":"An unprecedented number of viruses have been discovered by leveraging advances in high-throughput sequencing. Infectious clone technology is a universal approach that facilitates the study of biology and role in disease of viruses. In recent years homology-based cloning methods such as Gibson assembly have been used to generate virus infectious clones. We detail herein the preparation of home-made cloning materials for Gibson assembly. The home-made materials were used in one-step generation of the infectious cDNA clone of a plant RNA virus into a T-DNA binary vector. The clone was verified by a single Illumina reaction and a de novo read assembly approach that required no primer walking, custom primers or reference sequences. Clone infectivity was finally confirmed by <i>Agrobacterium</i>","dates":{"release":"2020-01-01T00:00:00Z","publication":"2020","modification":"2026-04-07T21:02:12.226Z","creation":"2022-02-09T08:12:26.632Z"},"accession":"S-EPMC7990137","cross_references":{"pubmed":["33768973"],"doi":["10.1186/s42483-020-00077-4"]}}