Recombination-assisted megaprimer (RAM) cloning
نویسندگان
چکیده
منابع مشابه
Recombination-assisted megaprimer (RAM) cloning
No molecular cloning technique is considered universally reliable, and many suffer from being too laborious, complex, or expensive. Restriction-free cloning is among the simplest, most rapid, and cost-effective methods, but does not always provide successful results. We modified this method to enhance its success rate through the use of exponential amplification coupled with homologous end-join...
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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...
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Cloning techniques are fundamental to molecular biology. Classically, recombinant plasmid and/or phage vectors are prepared in vitro, the transformation step only serving for amplification purposes. A different approach would exploit the natural intracellular enzymatic machinery to produce recombinant DNA molecules. Such techniques are well-known to mutagenize chromosomal DNA by e.g. transposon...
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The technique of site-directed mutagenesis (SDM) is widely used in the field of molecular biology to introduce mutations into DNA (5). The development of polymerase chain reaction (PCR)-based SDM has eliminated the need to clone DNA, therefore simplifying the technique and enabling the desired results to be achieved more quickly (3). The “megaprimer” method of PCR-based SDM incorporates three p...
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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: MethodsX
سال: 2014
ISSN: 2215-0161
DOI: 10.1016/j.mex.2014.05.001