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Assembling Multiple Fragments: The Gibson Assembly
Methods in Molecular Biology, 2023The Gibson Assembly is a popular method for molecular cloning which has been developed specifically to join several fragments together in a specific order, without the constraint of restriction enzyme sites. This method is based on the assembly of overlapping fragments, generally produced by PCR, and then combining them using three enzymes: a 5 ...
Luisana Avilan
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Gibson Assembly facilitates bacterial allelic exchange mutagenesis [PDF]
Journal of Microbiological Methods, 2018 Allelic exchange mutagenesis that relies on RecA-mediated homologous recombination up- and downstream from the targeted gene is a generalizable method of site-specific bacterial gene knock-out and knock-in. However, generation of a mutagenic DNA construct (alternative allele flanked by regions surrounding the gene target) and subsequent mutant ...
Oleksandra Rudenko, Andrew C Barnes
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DNA library construction using Gibson Assembly® [PDF]
Nature Methods, 2015 Since its introduction 6 years ago, the Gibson Assembly method has been widely adopted as a preferred cloning method. Here we describe an application of Gibson Assembly beyond routine cloning: assembly in DNA library construction. Advantages of using Gibson Assembly, specifically the SGI-DNA Gibson Assembly HiFi 1 Step kit, in library construction ...
Steven Thomas +2 more
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Preparing Genes for Repetitive Elastin-Like Polypeptides Using Gibson Assembly
Methods in Molecular Biology, 2018Elastin-like polypeptides (ELPs) are stimuli-responsive polymers consisting of peptide repeat units from the elastin protein. Most commonly these materials are biosynthesized in E. coli using genes that have been built by standard molecular biology techniques.
Nolan B Holland
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Construction of an agroinfectious clone of bean rugose mosaic virus using Gibson Assembly
Virus Genes, 2017Construction of agroinfectious viral clones usually requires many steps of cloning and sub-cloning and also a binary vector, which makes the process laborious, time-consuming, and frequently susceptible to some degree of plasmid instability. Nowadays, novel methods have been applied to the assembly of infectious viral clones, and here we have applied ...
Tatsuya Nagata +2 more
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Site-Directed Mutagenesis by Gibson Assembly
Genetic Engineering & Biotechnology News, 2013Ezra Schildkraut, Peichung Hsieh
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A Modified Gibson Assembly Method for Cloning Large DNA Fragments with High GC Contents
Methods in Molecular Biology, 2018Yinhua Lu, Weihong Jiang, Lei Li
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