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describes about principle, steps involved and applications of PFGE

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  1. 1. PULSE FIELD GEL ELECTROPHORESIS By Anjali Naik Asst. Professor Institute of Forensic Science, Aurangabad
  2. 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. 3. principle
  4. 4. Steps involved • Preparation of cell suspension • Generation of plugs containing cells • DNA isolation in plug itself • Restriction digestion in plug itself • PFGE
  5. 5. instrument
  6. 6. 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
  7. 7. limitations • Time cunsuming • Needs skilled persons • Links for viedos •