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Pfge

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Anjali Naik

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.

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PULSE FIELD GEL ELECTROPHORESIS By Anjali Naik Asst. Professor Institute of Forensic Science, Aurangabad
? 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¨C20 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
principle
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Steps involved ? Preparation of cell suspension ? Generation of plugs containing cells ? DNA isolation in plug itself ? Restriction digestion in plug itself ? PFGE
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instrument
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
limitations ? Time cunsuming ? Needs skilled persons ? Links for viedos ? https://youtu.be/__QhCX12h8I https://youtu.be/IXxx3bjjR0E

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Pfge

  • 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¨C20 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
  • 6. Steps involved ? Preparation of cell suspension ? Generation of plugs containing cells ? DNA isolation in plug itself ? Restriction digestion in plug itself ? PFGE
  • 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