ArticlePloS one2011
Quick and clean cloning: a ligation-independent cloning strategy for selective cloning of specific PCR products from non-specific mixes.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 44 citations in OpenAlex.
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- DAPE cloning with modified primers for producing designated lengths of 3' single-stranded ends in PCR products.PloS one · 2025Article
- Compositional and temporal division of labor modulates mixed sugar fermentation by an engineered yeast consortium.Nature communications · 2024Article
- In vitro functional genetic modification of canine adenovirus type 2 genome by CRISPR/Cas9.Laboratory investigation; a journal of technical methods and pathology · 2021Article
- Engineering Betalain Biosynthesis in Tomato for High Level Betanin Production in Fruits.Frontiers in plant science · 2021Article
- A double-Flp-in method for stable overexpression of two genes.Scientific reports · 2020Article
- Article
- Mutant T4 DNA polymerase for easy cloning and mutagenesis.PloS one · 2019Article
- Homologous alignment cloning: a rapid, flexible and highly efficient general molecular cloning method.PeerJ · 2018Article
- TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression.PloS one · 2017Article
- Systematic analysis of the kalimantacin assembly line NRPS module using an adapted targeted mutagenesis approach.MicrobiologyOpen · 2016Article
- Production of retroviral constructs for effective transfer and expression of T-cell receptor genes using Golden Gate cloning.BioTechniques · 2015Article
- Complementary annealing mediated by exonuclease: a method for seamless cloning and conditioning site-directed mutagenesis.Biotechnology, biotechnological equipment · 2015Article
- IRDL cloning: a one-tube, zero-background, easy-to-use, directional cloning method improves throughput in recombinant DNA preparation.PloS one · 2014Article
- Hot Fusion: an efficient method to clone multiple DNA fragments as well as inverted repeats without ligase.PloS one · 2014Article
- Generating in vivo cloning vectors for parallel cloning of large gene clusters by homologous recombination.PloS one · 2013Article
- Article
- Ultra-low background DNA cloning system.PloS one · 2013Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We have developed an efficient strategy for cloning of PCR products that contain an unknown region flanked by a known sequence. As with ligation-independent cloning, the strategy is based on homology between sequences present in both the vector and the insert. However, in contrast to ligation-independent cloning, the cloning vector has homology with only one of the two primers used for amplification of the insert. The other side of the linearized cloning vector has homology with a sequence present in the insert, but nested and non-overlapping with the gene-specific primer used for amplification. Since only specific products contain this sequence, but none of the non-specific products, only specific products can be cloned. Cloning is performed using a one-step reaction that only requires incubation for 10 minutes at room temperature in the presence of T4 DNA polymerase to generate single-stranded extensions at the ends of the vector and insert. The reaction mix is then directly transformed into E. coli where the annealed vector-insert complex is repaired and ligated. We have tested this method, which we call quick and clean cloning (QC cloning), for cloning of the variable regions of immunoglobulins expressed in non-Hodgkin lymphoma tumor samples. This method can also be applied to identify the flanking sequence of DNA elements such as T-DNA or transposon insertions, or be used for cloning of any PCR product with high specificity.
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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.