ArticleThoracic cancer2020
Cloning short DNA into plasmids by one-step PCR.
Article in Thoracic cancer, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 8 citations in OpenAlex.
- A 3D Printed Mini-Gel Electrophoresis System for Rapid and Inexpensive DNA Nanoswitch Biosensing.ACS measurement science au · 2026Article
- A 3D printed mini-gel electrophoresis system for rapid and inexpensive DNA nanoswitch biosensing.bioRxiv : the preprint server for biology · 2026Article
- Improved vectors for retron-mediated CRISPR-Cas9 genome editing in Saccharomyces cerevisiae.G3 (Bethesda, Md.) · 2025Article
- Pepper (Capsicum annuum L.) AP2/ERF transcription factor, CaERF2 enhances salt stress tolerance through ROS scavenging.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2025Article
- TA-Cloning for Diabetes Treatment: Expressing Corynebacterium Malic Enzyme Gene in E. coli.Current microbiology · 2024Article
- Unraveling the functional consequences of a novel germline missense mutation (R38C) in the yeast model of succinate dehydrogenase subunit B: insights into neurodegenerative disorders.Frontiers in molecular neuroscience · 2023Article
Corrections and comments
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Authors and funding
8 authors at 4 institutions in 1 country.
Funding
Abstract
backgroundPlasmid construction of small fragments of interest (such as insertion of small fragment marker genes, expression of shRNA, siRNA, etc) is the basis of many biomolecular experiments. Here, we describe a method to clone short DNA into vectors by polymerase chain reaction (PCR), named one-step PCR cloning. Our method uses PCR to amplify the entire circular plasmid. The PCR was performed by the primers containing the gene of short DNA with overlapping sequences between 10-15 bp. The PCR products were then transformed into E. coli and cyclized by homologous recombination in vivo.
methodsThe pEGFP-N1-HA plasmid was constructed by one-step PCR and transformation. Cells were transfected with pEGFP-N1-HA and pEGFP-N1 plasmid using TurboFect transfection reagent. Protein expression was detected by western blotting and the HA-GFP fusion protein was detected by confocal microscopy.
resultsThe pEGFP-N1-HA plasmid was successfully constructed and HA expression in cells.
conclusionsFree from the limitations of restriction enzyme sites and omitting the ligation process, our method offers a flexible and economical option of plasmid construction. KEY POINTS: Significant findings of the study A method to clone short DNA into plasmids was found. What this study adds Our study provides a flexible and economical option to clone short DNA into plasmids.
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Registered trials
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.