MCT cloning: a seamless cloning strategy for inserting DNA fragments
نویسندگان
چکیده
منابع مشابه
A strategy for seamless cloning of large DNA fragments from Streptomyces.
We report a novel method for the seamless cloning of large DNA fragments (SCLF) of up to 44 kb or larger from Streptomyces chromosomal DNA. SCLF is based on homologous recombination in Streptomyces and is easy to perform. The strategy of SCLF is to flank the target sequence in the chromosomal DNA with two identical restriction sites by the insertion of plasmids containing that site at either en...
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Here, an efficient cloning strategy for large DNA fragments and for simultaneous assembly of multiple DNA fragments assembly is presented. This strategy is named OEPR (based on Overlap Extension PCR and Recombination in vivo). OEPR cloning is a seamless, restriction- and ligation-independent method. The method takes advantage of both homologous recombination enzymes in E. coli and overlap PCR. ...
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Recombinant DNA vectors are fundamental tools in molecular biology and genetic engineering. One of the problems frequently encountered in vector construction is the lack of compatible restriction sites between the vectors and the DNA inserts. The conventional approach to solve this problem is to make the DNA fragments blunt-ended (3,4,12) or to add linkers or adaptors (10,11,14) before ligation...
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An effective technique using an Escherichia coli plasmid system was developed to clone fragments of exogenous DNA of as large as 100 kilobase pairs. The characteristic features of this technique are the use of a low copy number (one to two) mini-F based plasmid vector and the introduction of artificial lambda cosR ends into the termini of DNA sources and then of the cosL ends into those of line...
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Background & objective: Tuberculosis (TB) remains a major cause of death around the world. Bacillus Calmette Guérin (BCG) is the only vaccine used in TB prevention that has a protective effect in children, but its effectiveness declines in adults. Design and development of new vaccines is the most effective way against TB. The aim of this study was to design and construc...
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ژورنال
عنوان ژورنال: Biotechnology & Biotechnological Equipment
سال: 2018
ISSN: 1310-2818,1314-3530
DOI: 10.1080/13102818.2018.1507756