ArticleBiotechnology, biotechnological equipment2015
Complementary annealing mediated by exonuclease: a method for seamless cloning and conditioning site-directed mutagenesis.
Article in Biotechnology, biotechnological equipment, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- SECISBP2L-Mediated Selenoprotein Synthesis Is Essential for Autonomous Regulation of Oligodendrocyte Differentiation.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2022Article
- An improved overlap extension PCR for simultaneous multiple sites large fragments insertion, deletion and substitution.Scientific reports · 2019Article
- TT(N)mGCCTC inhibits archaeal family B DNA polymerases.Scientific reports · 2018Article
- Characterization of a cytoplasmic glucosyltransferase that extends the core trisaccharide of theThe Journal of biological chemistry · 2017Article
- Strategies used for genetically modifying bacterial genome: site-directed mutagenesis, gene inactivation, and gene over-expression.Journal of Zhejiang University. Science. B · 2016Review
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Authors and funding
5 authors.
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Abstract
Traditional cut-paste DNA cloning is often limited by the availability of restriction enzyme sites. Here, we described the complementary annealing mediated by exonuclease (CAME), in which the insert or vector fragment is amplified to carry sequences complementary to the other, and both fragments are modified by exonuleases to create directional single-stranded overhangs. The two recessed DNA fragments are joined through complementary strand annealing. The CAME is highly efficient for cloning the DNA of at least 12 kb and single DNA fragment out of a complex DNA sample. Moreover, the application of CAME greatly improved the efficiency of site-directed mutagenesis.
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