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For their use as vectors, and for molecular cloning , plasmids often need to be isolated.

The yield is a small amount of impure plasmid DNA, which is sufficient for analysis by restriction digest and for some cloning techniques. In the latter, much larger volumes of bacterial suspension are grown from which a maxi-prep can be performed. In essence, this is a scaled-up miniprep followed by additional purification. This results in relatively large amounts several hundreds micrograms of very pure plasmid DNA. In recent times, many commercial kits have been created to perform plasmid extraction at various scales, purity, and levels of automation.

Plasmid DNA may appear in one of five conformations, which for a given size run at different speeds in a gel during electrophoresis. The conformations are listed below in order of electrophoretic mobility speed for a given applied voltage from slowest to fastest:. The rate of migration for small linear fragments is directly proportional to the voltage applied at low voltages.

At higher voltages, larger fragments migrate at continuously increasing yet different rates. Thus, the resolution of a gel decreases with increased voltage. At a specified, low voltage, the migration rate of small linear DNA fragments is a function of their length. Large linear fragments over 20 kb or so migrate at a certain fixed rate regardless of length.

This is because the molecules 'resperate', with the bulk of the molecule following the leading end through the gel matrix. Restriction digests are frequently used to analyse purified plasmids. These enzymes specifically break the DNA at certain short sequences. The resulting linear fragments form 'bands' after gel electrophoresis. It is possible to purify certain fragments by cutting the bands out of the gel and dissolving the gel to release the DNA fragments.

The use of plasmids as a technique in molecular biology is supported by bioinformatics software. These programs record the DNA sequence of plasmid vectors, help to predict cut sites of restriction enzymes , and to plan manipulations. These software help conduct entire experiments in silico before doing wet experiments. One can find and request plasmids from those databases for research. Researcher also often upload plasmid sequences to the NCBI database , from which sequences of specific plasmids can be retrieved.

From Wikipedia, the free encyclopedia. Further information: Vector molecular biology. Main article: Cloning vector. Main article: Expression vector. Main article: Vectors in gene therapy. Main article: Addiction module. Acta Microbiologica et Immunologica Hungarica.

Transformation of Agrobacterium Tumefactions by Conjugation Technique

Microbiology and Molecular Biology Reviews. Bacterial Plasmids. Encyclopedia of Life Sciences. In Georg Lipps eds. Plasmids: Current Research and Future Trends. Caister Academic Press. Physiological Reviews.

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In Nicola Casali, Andrew Presto eds. Coli Plasmid Vectors: Methods and Applications. Methods in Molecular Biology. Humana Press. Microbiology Society.

Molecular Biology, Pathogenicity, and Ecology of Bacterial Plasmids -

Brown The Biology of Plasmids. Wiley-Blackwell; First Edition. Clark; Nanette Jean Pazdernik Molecular Biology 2nd ed. Academic Cell. Smith and R. Elizabeth Sockett, eds.

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Y: Cold Spring Harbor Laboratory. Applied Microbiology and Biotechnology. Trends in Biotechnology. Streips, Ronald E. Yasbin, eds. Modern Microbial Genetics 2nd ed. In Nicola Casali, Andrew Preston eds. Methods in Molecular Biology, Vol. The notion of the episome" PDF. Journal of Biosciences.

3.1: Horizontal Gene Transfer in Bacteria

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Molecular Biology, Pathogenicity, and Ecology of Bacterial Plasmids

Archived from the original PDF on 24 July Bibcode : PNAS Microbial Biotechnology. Journal of Bacteriology. The DNA Lab.

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