METHOD FOR RAPID TYPIFICATION OF MICROORGANISMS BY PULSE FIELD GEL ELECTROPHORESIS (PFGE) WHEREBY THE SAMPLES ARE PREPARED IN AN AUTOCLAVE AND FLEXIBLE MOULD

摘要

Disclosed is a method for rapidly typifying yeasts, parasites and bacteria.The method involves the following steps: a) Preparing intact and immobilizedDNA within 5 to 60 minutes by means of a method which uses a set of reagentsthat only contains a buffer solution, a detergent, a chelating agent and anagent that breaks up the hydrogen bridges. b) Separating intact DNA moleculesor their restriction fragments using pulsed field electrophoresis mini-equipment of systems CHEF (Contour Clamped Homogeneous Electric Field) andTAFE (Transversal Alternating Field Electrophoresis) for time periods rangingbetween 2.5 and 7 hours. c) Selecting the optimal conditions that will beapplied in the miniCHEF by using a method simulating a priori theelectrophoretic patterns that would be obtained in said gels. d) Providing thereorientation times, migration speeds and molecule sizes without using sizemarkers but using a method that analyzes migrated distances.