Analysis of Nucleic Acids by Capillary Electrophoresis by Christoph Heller (auth.), Christoph Heller (eds.)

By Christoph Heller (auth.), Christoph Heller (eds.)

During the decade, capillary electrophoresis has been constructed right into a very robust analytical process, which has many merits over traditional slab gel ideas. the advance is similar to the single occuring previous within the box of chromatography, with the advent of the excessive functionality know-how. How very important this system has be­ come, is mirrored by way of the shear quantity of papers released every month; in addition to a dozen of books already released in this topic. essentially the most vital meetings within the box, the "International Symposium on excessive functionality Capillary Electrophoresis" draws now approximately thousand humans each year. As capillary electrophoresis could be utilized to many alternative analytical difficulties, a spe­ cialization is unavoidable. This evolution can also be mirrored within the improvement of instrumen­ tation: while the 1st units have been designed for all attainable purposes, new tools are actually equipped, which are really good for one specific job, e.g. DNA research. I greatly welcome the choice of the sequence editor and the writer, to edit a sequence ofspecialized books, overlaying all elements of capillary electrophoresis. Having labored at the electrophoretic separation of DNA for a few years, i'm confident that there are such a lot of various points in this factor that they deserve an entire booklet on their lonesome. for that reason, i used to be chuffed to agree while being requested to edit this book.

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This, in effect, reduces the problem of the dynamics of the solute inside the gel, which can be quite involved, to a much simpler (static) geometrical one . Using Ogston's result, we then find that rt(R+r)2 l! * = f V = e" KC = e 4aC 2 '" e (17) where a (C) - C - 112 is the average pore size as derived by Ogston and K - (R + r)2 is the retardation coefficient. The last part ofEq . (17) applies to flexible DNAs. This expression is intuitively' attractive; indeed, the mobility decreases exponentially with the square of the ratio of the effective particle size R + r to the mean pore size a (C), and goes rapidly to zero when (R + r) > a or M o > M a .

Smith and B. L. Karger, "Rapid separation of DNA restriction fragments using capillary electrophoresis", J. Chromatography, 4581988323-333 . [35] T. J. Kasper, M. Melera, P. Gozel and R. G. Brownlee, "Separation and detection of DNA by capillary electrophoresis", J. Chromatography, 458 1988 303-312. [36] M. Zhu , D. L. Hansen, S. Burd and F. Gannon , "Factors affecting free zone electrophoresis and isoelectric focusing in capillary electrophoresis", J. Chromatography, 480 1989 311-319. [37] M. Strege and A.

The particle radius R : the radius r is given by the negat ive extrapolated value of R for which K(R) == 0 [53-55] . Again, R is given here by the radius of gyration Rg(M) of the DNA mole cule. Curiously, although this model is based on Ogston's calculations, which predict a- l/CIl2 , this scaling law is rarely observed when the estimated mean pore size is plotted vs. the gel concentration C [II , 54] . For example, we found a - CO·65 in ref. [55] . This discrepancy suggests that there is a problem with this geometrical model, or perhaps with its interpretation.

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