DNA molecules in an agarose matrix elongate and migrate toward the anode when exposed to an electric field. An agarose matrix represents a highly irregular network with pores of various dimensions, large open areas, and regions with different densities. During electrophoresis, migration will be retarded by obstructions that prevent sieving by size and result in "end-on" migration as if through ...

MATERIALS 5x TBE electrophoresis buffer Polyacrylamide gels are poured and run in 0.5x or 1x TBE at low voltage (1-8 V/cm) to prevent denaturation of small fragments of DNA by Joulic heating. Other electrophoresis buffers such as 1x TAE (please see Agarose Gel Electrophoresis) can be used, but they are not as good as TBE. The gel must be run more slowly in 1x TAE, which does not provide as much...

INTRODUCTION A bottleneck for high-throughput proteomic studies is image analysis. In conventional two-dimensional (2D) methodology, protein samples are separated on individual gels, stained, and quantified, followed by image comparison with computer-aided image analysis programs. Since different images are not always perfectly superimposable, image analysis is very time consuming. Fluorescent ...

background and objectives: citrobacter freundii is an opportunistic pathogen causing nosocomial infections and resistant to various antibiotics. we aimed to determine the clonal relationship of c. freundii isolates using pulsed-field gel electrophoresis (pfge). materials and methods: fifty clinical isolates of c. freundii were collected from the main hospital in kermanshah. after antibiotic sus...

2,2,2-Trichloroethanol (TCE) incorporated into polyacrylamide gels before polymerization provides fluorescent visible detection of proteins in less than 5min of total processing time. The tryptophans in proteins undergo an ultraviolet light-induced reaction with trihalocompounds to produce fluorescence in the visible range so that the protein bands can be visualized on a 300-nm transilluminator...

Anion transport inhibitors, such as SITS (4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid) and heparin, inhibit reversibly the bicarbonate-sensitive adenylylcyclase of porcine sperm plasma membrane. In the light of this, SITS- and heparin-affinity chromatographies were applied in order to purify sperm adenylylcyclase. SITS-Affi-Gel 102 binds proteins extracted from the porcine cauda ep...

Conformation-sensitive gel electrophoresis is a useful method for identifying allele polymorphism; it provides co-dominant molecular markers. Using this method, we identified genetic variability in the third intron of the fibroin light chain gene, fib-L, in six Bombyx mori strains. Only Chinese C21A strain did not demonstrate allelic alterations, showing only homoduplex DNA molecules. We found ...