General Method for Plasmid Construction Using Homologous Recombination
نویسندگان
چکیده
منابع مشابه
General method for plasmid construction using homologous recombination.
We describe a general method for plasmid assembly that uses yeast and extends beyond yeast-specific research applications. This technology exploits the homologous recombination, double-stranded break repair pathway in Saccharomyces cerevisiae to join DNA fragments. Synthetic, double-stranded "recombination linkers" were used to "subclone" a DNA fragment into a plasmid with > 80% efficiency. Qua...
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The cloning of DNA fragments is a fundamental aspect of molecular biology. Traditional DNA cloning techniques rely on the ligation of an insert and a linearized plasmid that have been digested with restriction enzymes and the subsequent introduction of the ligated DNA into Escherichia coli for propagation. However, this method is limited by the availability of restriction sites, which often bec...
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Vol. 35, No. 4 (2003) BioTechniques 693 points are distributed along lines that represent the expected differences in signal intensity from each channel. Without a systematic dye effect or after proper normalization, we would see three sets of data points along three equally spaced lines parallel with slope one. On the other hand, the direct comparison of measurements for each dye on different ...
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The levels of intramolecular plasmid recombination, following transfection of a plasmid substrate for homologous recombination into normal and immortally transformed cells, have been examined by two independent assays. In the first assay, recovered plasmid was tested for DNA rearrangements which regenerate a functional neomycin resistance gene from two overlapping fragments. Following transform...
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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...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 1999
ISSN: 0736-6205,1940-9818
DOI: 10.2144/99261rr02