DNA library construction using Gibson Assembly®

Gibson Assembly® Primer-Bridge End Joining (PBnJTM) Cloning Gibson Assembly® Cloning without homologous overlap regions

Gibson Assembly® cloning has proven to be useful as a molecular biology technique for the seamless assembly of synthetic and natural genes and large-scale genetic pathways. Notably, Gibson Assembly cloning has enabled the synthesis of the first bacterial genome1, the first synthetic cell2, and the first minimal cell3. Additionally, the Gibson Assembly method has been utilized for genetic recodi...

A Simplified and Versatile System for the Simultaneous Expression of Multiple siRNAs in Mammalian Cells Using Gibson DNA Assembly

RNA interference (RNAi) denotes sequence-specific mRNA degradation induced by short interfering double-stranded RNA (siRNA) and has become a revolutionary tool for functional annotation of mammalian genes, as well as for development of novel therapeutics. The practical applications of RNAi are usually achieved by expressing short hairpin RNAs (shRNAs) or siRNAs in cells. However, a major techni...

Construction of a genomic DNA library by TA cloning.

Transient gene expression in tobacco using Gibson assembly and the Gene Gun.

In order to target a single protein to multiple subcellular organelles, plants typically duplicate the relevant genes, and express each gene separately using complex regulatory strategies including differential promoters and/or signal sequences. Metabolic engineers and synthetic biologists interested in targeting enzymes to a particular organelle are faced with a challenge: For a protein that i...

Preparation of single rice chromosome for construction of a DNA library using a laser microbeam trap.

We report the development of a laser micromanipulation system and its application in the isolation of individual rice chromosomes directly from a metaphase cell. Microdissection and flow sorting are two major methods for the isolation of single chromosome. These methods are dependent on the techniques of chromosome spread and chromosome suspension, respectively. In the development of this syste...