Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction.

By R. Horton, Z. Cai, S. Ho, L. Pease. BioTechniques, 2013

1,384 citations recorded by the source corpus.

Gene Splicing by Overlap Extension or "gene SOEing" is a PCR-based method of recombining DNA sequences without reliance on restriction sites and of directly generating mutated DNA fragments in vitro. By modifying the sequences incorporated into the 5'-ends of the primers, any pair of polymerase chain reaction products can be made to share a common sequence at one end. Under polymerase chain reaction conditions, the common sequence allows strands from two different fragments to hybridize to one another, forming an overlap. Extension of this overlap by DNA polymerase yields a recombinant molecule. This powerful and technically simple approach offers many advantages over conventional approaches for manipulating gene sequences.

Subfields: Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/01a636966924a8b55e8e00cb30f3823a7dba948a

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Citations in the corpus

In vivo recombineering of bacteriophage λ by PCR fragments and single-strand oligonucleotides (2004, methodology): “(v) Using appropriate PCR primers and the gene SOEing technique (Horton et al., 1990), we created a linear DNA product containing an intact copy of the wild-type E P gene adjoining a precise deletion of the entire ren gene but with homology beyond ren in the ninR region of the phage.”
Enzymatic Assembly of Overlapping DNA Fragments (2011, methodology): “Overlapping DNA fragments can be assembled during PCR (Horton et al., 1990) using primers specific to the ends of the assembly; however, the size of the product cannot exceed the length that can be readily amplified by PCR. Complementary vector sequence is added to PCR primers in a technique known…”
BglBricks: A flexible standard for biological part assembly (2010, methodology): “To name a few, Gene Splicing by Overlap Extension (gene SOEing), is a sequence-independent, PCR-based method, that enables site-directed mutagenesis and/or recombination of DNA sequences without relying on restriction enzyme sites [25,26].”
Synthetically-primed adaptation of Pseudomonas putida to a non-native substrate D-xylose (2024, neutral): “In differential abundance analysis, 4 post-FMT species were signi ﬁ cantly more abundant in patients wholaterdeveloped gradeIII – IVaGVHD vs. those who did not (Fig.”