Pulse field gel electrophoresis is a technique used to separate high molecular weight DNA fragments. It involves embedding DNA in agarose plugs, digesting the DNA within the plugs using restriction enzymes, and then applying an alternating electric field to separate DNA fragments of varying sizes. Pulse field gel electrophoresis is commonly used for bacterial strain typing in epidemiological studies and provides a gold standard for linking food/environmental isolates to clinical infections. However, it is time-consuming and requires skilled technicians to perform.

1. PULSE FIELD GEL
ELECTROPHORESIS
By
Anjali Naik
Asst. Professor
Institute of Forensic Science, Aurangabad

2. • Variation of gel electrophoresis used for
separation of high molecular weight DNA
• High molecular weight DNA can not separate
properly by routine gel electrophoresis
• DNA molecules larger than 15–20 kb migrating
through a gel will essentially move together in a
size independent manner.
• It result in the formation of single band after
electrophoresis consisting of fragments of various
sizes
• At Columbia University in 1984, David C. Schwartz
and Charles Cantor developed PFGE

3. principle

6. Steps involved
• Preparation of cell suspension
• Generation of plugs containing cells
• DNA isolation in plug itself
• Restriction digestion in plug itself
• PFGE

10. instrument

11. Applications
• PFGE may be used for genotyping or genetic
fingerprinting.
• It is commonly considered a gold standard in
epidemiological studies of pathogenic
organisms.
• Subtyping has made it easier to discriminate
among strains of Listeria monocytogenes and
thus to link environmental or food isolates
with clinical infections

12. limitations
• Time cunsuming
• Needs skilled persons
• Links for viedos
• https://youtu.be/__QhCX12h8I
https://youtu.be/IXxx3bjjR0E