What is the principle of pulsed field gel electrophoresis?

PFGE resolves DNA by alternating the electrical field between spatially distinct pairs of electrodes. This technique results in the separation of DNA fragments of up to ~10 Mb by their reorientation and movement at different speeds through the pores of an agarose gel.

What is pulsed field gel electrophoresis Neb?

Pulsed-field gel electrophoresis (PFGE) is a laboratory technique used by scientists to produce a DNA fingerprint for a bacterial isolate. A bacterial isolate is a group of the same type of bacteria. PulseNet investigates bacterial isolates from sick people, contaminated food, and the places where food is produced.

What are the applications of pulsed field gel electrophoresis?

PFGE in microbiology is a standard method which is used for typing of bacteria. It is also a very useful tool in epidemiological studies and gene mapping in microbes and mammalian cell, also motivated development of large-insert cloning system such as bacterial and yeast artifical chromosomes.

When and why the pulse field gel electrophoresis is preferred?

Pulsed-field gels are more effective than regular gels at achieving differential separation of the small and large DNA molecules due to the constant change in the direction of the electric field in the gel. Pulsed-field gel electrophoresis can resolve DNA out to 100 kb and beyond.

What are 5 kinds of pulsed-field gel electrophoresis?

Different types of PFGE units are currently available including contour-clamped homogeneous electric field (CHEF) [71,72], transverse alternating field electrophoresis (TAFE) [73] and its relative ST/RIDEtm (Stratagene), and rotating gel electrophoresis (RGE) [74,75], with CHEF being the most common one.

How long does pulsed-field gel electrophoresis?

PFGE takes 2–3 days, excluding sample preparation.

What is the duration of the pulse used during pulse field electrophoresis?

Electrophoresis is performed with a horizontal gel electrophoresis apparatus and a suitable pulse controller to vary current direction. We run gels under thermostatically controlled conditions (14°) in a Bio-Rad CHEF system, using the following parameters: 200 V for 20 hr with a pulse ramp of 4 to 40 sec.