What is a PFGE pattern?

The fingerprints that scientists use to identify bacteria are called PFGE patterns. PFGE stands for Pulsed-Field Gel Electrophoresis. Scientists find bacterial fingerprints by cutting the bacteria’s DNA into tiny pieces and then placing them on a gel, which is a flat slab of gelatin.

What is PFGE and how does it work?

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 is the main limitation of PFGE?

The main disadvantage of PFGE is that it is labour-intensive with up to 3 to 4 days required to complete the lengthy PFGE protocol, and it has a high chance of yielding inconclusive results when compared to AFLP (Klaassen et al. 2002).

How does PFGE separate larger fragments?

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.

Is PFGE still used?

For over 20 years, PulseNet has used a DNA fingerprinting technology called PFGE to detect and help solve foodborne disease outbreaks. Using PFGE, PulseNet prevents an estimated 270,000 illnesses and saves at least half a billion dollars in medical costs and lost productivity each year.

How does PFGE separate DNA with different structures?

Pulsed-field gel electrophoresis (PFGE) is a method widely used to separate fragments of DNA as long as several million bases by subjecting the gel to an electrical current alternately delivered from two angles in timed intervals, which minimizes diffusion of large molecules.

Why PFGE is considered as gold standard in pathogen typing?

Pulsed-field gel electrophoresis (PFGE) is considered the “gold standard” for bacteria typing. The method involves enzyme restriction of bacteria DNA, separation of the restricted DNA bands using a pulsed-field electrophoresis chamber, followed by clonal assignment of bacteria based on PFGE banding patterns.

Which of the following is are applications of PFGE?

In addition to the genotyping and fingerprinting, other applications of PFGE include the use of it in aetiologic studies of bacteria, identification of bacteria isolated from environmental or clinical samples, identification of antibiotics resistant strains (e.g. MRSA strains) as well as the classification (taxonomy) …

Why are cells put in plugs in PFGE?

PFGE involves the isolation of the intact chromosomal DNA by lysing bacterial cells embedded in an agarose plug to avoid the mechanical shearing of DNA molecules during the extraction (21).

How much does PFGE cost?

around USD$30,000
PFGE equipment is expensive and exclusively sold by two providers [Bio Rad, Analytik Jena], they cost around USD$30,000 and do not have an open source version.

Does SDS denature DNA?

These additives keep the solutes or molecules in a denatured state during separation. Urea denatures double-stranded DNA to single-stranded DNA. A detergent, sodium dodecyl sulfate (SDS), denatures proteins.