Question

1- what is advantage does pulsed field gradient electrophoresis in gels have over conventional slab gel electrophoresis ? 2- what samples uniquely are suited for this technique and why? 3- discuss the motivation and resistance to adopting new , faster methods of gel electrophoresis that are in capillaries and not on slabs? please give me also refrences

Answer 1

1. pulsed-field gel electrophoresis (PFGE) has huge advantages for molecular biology research than conventional slab gel electrophoresis. Standard gel electrophoresis was unable to separate very large molecules of DNA effectively. DNA molecules larger than 15-20 kb migrating through a gel will essentially move together in a size-independent manner. The development of PFGE expanded the range of resolution for DNA fragments by as much as two orders of magnitude.

1. pulsed-field gel electrophoresis (PFGE) separates DNA molecules up to 2000 kb with resolutions exceeding the logarithmic molecular weight dependence of conventional electrophoresis.

1. Electrophoresis is a separation techniques it will separate analytes by their ability to move through a conductive medium, usually an aqueous buffer, in response to an applied electric field.

In slab gel electrophoresis the conducting buffer is retained within a porous gel of agarose or polyacrylamide. Slabs are formed by pouring the gel between two glass plates separated by spacers.

In capillary electrophoresis, the conducting buffer is retained within a capillary tube whose inner diameter is typically 25–75 μm. Samples are injected into one end of the capillary tube. As the sample migrates through the capillary its components separate and elute from the column at different times

Reference: Schwartz, David C., and Charles R. Cantor. "Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis." Cell 37.1 (1984): 67-75.