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 used for?

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.

How are the large fragments of DNA protected for PFGE separation?

This is done to protect the chromosomal DNA from mechanical damage by immobilizing it into agarose blocks. Then the bacterial cells are lysed to release the DNA. The agarose-DNA suspension is also known as plug mold.

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).

Which is the best fingerprinting method for PulseNet?

PFGE is the current “gold standard” fingerprinting method used within PulseNet. However, PulseNet is transitioning toward using whole genome sequencing (WGS). For a few organisms, PulseNet also uses multi-locus variable tandem repeat analysis (MLVA) to aid outbreak investigations. PFGE Process: Larger View.

How does pulsed field gel electrophoresis ( PFGE ) 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.

What are the advantages and disadvantages of PFGE?

Advantages of PFGE High concordance with epidemiological relatedness Can be applied as a universal generic subtyping method for many different bacteria with only the choice of the restriction enzyme and electrophoresis conditions optimized for each species Stable and reproducible DNA restriction patterns

How is PFGE different from conventional DNA electrophoresis?

PFGE is different from conventional DNA electrophoresis because PFGE can separate very large fragments to generate a fingerprint by constantly changing the direction of the electric field.