Controlling electrophoretic trapping of circular DNA by addition of starch preparations to agarose gels

Starch preparations were added to agarose gels to enhance the electrophoretic trapping of circular plasmid DNA. The critical voltages required to trap the open circular (OC) and the supercoiled (SC) forms of a 13.1-kbp plasmid were measured in gels composed of agarose and added starch preparations....

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Bibliographic Details
Published in:Applied biochemistry and biotechnology 2001-07, Vol.95 (1), p.31-43
Main Authors: Cole, K.D, Tellez, C.M, Nguyen, R
Format: Article
Language:English
Subjects:
agarose
amylose
Amylose - chemistry
Biochemistry
Biological and medical sciences
Biotechnology
Carbohydrates
Deoxyribonucleic acid
DNA
DNA, Circular - chemistry
DNA, Circular - isolation & purification
DNA, Superhelical - chemistry
DNA, Superhelical - isolation & purification
electric power
Electrophoresis, Agar Gel - methods
Fundamental and applied biological sciences. Psychology
gel electrophoresis
Gels
Methods. Procedures. Technologies
modified starch
Others
plasmids
Plasmids - genetics
Starch
Starch - chemistry
Studies
Trapping
Various methods and equipments
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Summary:Starch preparations were added to agarose gels to enhance the electrophoretic trapping of circular plasmid DNA. The critical voltages required to trap the open circular (OC) and the supercoiled (SC) forms of a 13.1-kbp plasmid were measured in gels composed of agarose and added starch preparations. Modified starch preparations reduced the critical voltage required to trap the OC form of the plasmid to approximately one-third of the control value (in 1% agarose gels). Amylose (a fraction of starch with a low amount of branching) also reduced the critical voltage to trap the OC form in a similar manner. The critical voltage to trap the SC form of the plasmid was not significantly reduced by the starch preparations. The capacity to trap OC DNA was increased by the addition of higher amounts of the starch preparations added to the gels. Field inversion gel electrophoresis was used to characterize the length of the traps in the gels. The starch preparations and amylose increased the trap lengths approximately twofold. The increased trap length correlated with the decreased critical voltage required to trap the OC form of the 13.1-kbp plasmid.
ISSN:0273-2289
1559-0291
0273-2289
DOI:10.1385/ABAB:95:1:31