High‐performance field inversion capillary electrophoresis of 0.1–23 kbp DNA fragments with low‐gelling, replaceable agarose gels

Abstract

Field inversion capillary gel electrophoresis (FICGE) has been used for the separation of 0.1-23 kbp DNA fragments in a low-melting, low-gelling agarose gel. The influence of the amplitude of the voltage pulses, the pulse times and gel concentrations on the separation factor has been studied and found to be similar to that of polyacrylamide gels. Very high resolution can be obtained by altering continuously the pulse times and/or voltage according to a program tailor-made for the size of the DNA molecules to be separated (programmed FICGE). The advantage of these agarose gels in comparison with polyacrylamide gels is that they are nontoxic, easy to prepare and they have high UV transmission. They give a resolution that is equivalent to or better than that of polyacrylamide gels, and the risk of bubble formation in the gel is small. In addition, they are replaceable and, therefore, very convenient for automated analyses.

Quaternary ammonium substituted agarose as surface coating for capillary electrophoresis

“…proteins. In CE, agarose has been used as electrochromatographic media 11 and sieving matrix both in the gel state 12,13 and in the sol state. 14 To the best of our knowledge there is, however, only one report on the use of agarose as surface coating in CE.…”

Electrophoretic Separation of Small DNA Fragments in the Presence of Electroosmotic Flow Using Poly(ethylene oxide) Solutions

“…Since the first report of DNA separation with cross-linked PA, several different polymers have been successfully employed in the analysis of DNA. − Currently, solutions prepared from linear polymers, such as PA, poly(ethylene oxide) (PEO), − and celluloses, , are most commonly used for high-speed and high-resolution DNA separation. The separation of larger DNA fragments can also be achieved even in very dilute (unentangled) polymer solutions, where the pore-based separation model is unsuitable …”

Identifying the Orientation of DNA Fragment in Recombinant Plasmid by Capillary Electrophoresis with a Non-Gel Sieving Solution

“…There are many factors that govern the electrophoretic separation quality. Because such a topic has been detailed by others, [14][15][16][17][18][19][20][21][22]25,26 only a brief description is given here.…”

“…Recently, linear polymers have been popularly used in CE as the sieving matrix instead of the crosslinked gel, in terms of non-gel sieving or an entangled polymersolution CE, 13,14 due to its facilitation 15 of high separation speed, long read length and replaceable operation compared to CGE. Many polymers have proved to be useful, including linear polyacrylamide and its derivatives, 14,16,17 cellulose and its derivatives, 18,19 polyethene oxide 20,21 and agarose 22 etc. Currently, non-gel sieving CE is being extensively applied in DNA sequencing, characterizing the polymorphism of DNA, analyzing PCR products, gene diagnosis and therapy etc.…”