BACKGROUND Somatic cell nuclear transfer (SCNT) using genetically engineered donor cells is currently the most widely used strategy to generate tailored pig models for biomedical research. Although this approach facilitates a similar spectrum of genetic modifications as in rodent models, the outcome in terms of live cloned piglets is quite variable. In this study, we aimed at a comprehensive an...

Production of recombinant proteins is the starting point for biochemical and biophysical analyses and requires methodology to efficiently proceed from gene sequence to purified protein. While optimized strategies for the efficient cloning of single-gene fragments for bacterial expression is available, efficient multiple DNA fragment cloning still presents a challenge. To facilitate this step, w...

background: the homologous recombination (hr) is one of the conventional cloning methods for the production of recombinant dna. it is a quick method for in vivo dna cloning without using the high cost restriction enzymes. a few modifications in the cloning procedure can increase the efficiency of this method. m aterials and methods: in this study, effect of heating on the rate of the igg1 heavy...

ed here improves upon the concept of directional cloning without forfeiting any of its advantages. The advantages include single-step cloning, high-efficiency generation of the desired recombinant molecule and low background self-ligation of vector. The added benefits of the triple-ligation strategy include easy resolution of vector fragments for gel purification, the ability to invert an inser...

BACKGROUND Polymerase chain reaction (PCR) is extensively applied in gene cloning. But due to the existence of introns, low copy number of particular genes and high complexity of the eukaryotic genome, it is usually impossible to amplify and clone a gene as a full-length sequence directly from the genome by ordinary PCR based techniques. Cloning of cDNA instead of genomic DNA involves multiple ...

The generation of DNA constructs is often a rate-limiting step in conducting biological experiments. Recombination cloning of single DNA fragments using the Gateway system provided an advance over traditional restriction enzyme cloning due to increases in efficiency and reliability. Here we introduce a series of entry clones and a destination vector for use in two, three, and four fragment Gate...