Abstract: Methods of use, accessories and chambers, optimal for performing Pulsed Field Gel Electrophoresis (PFGE) of DNA molecules in ‘Contour Clamped Homogeneous Electric Field’ (CHEF) and ‘Transversal Alternating Field Electrophoresis’ (TAFE) systems, are provided herein. DNA molecules are rapidly separated in the minigels of these chambers. The sizes of chambers and accessories are determined by the separation between the opposite polarity electrodes; which is comprised between 6.2 and 15 cm. Reproducibility of molecule separation is achieved because the accessories warrant homogeneous electric resistance in the buffer and minigels. Chambers allow a high-throughput sample format using the reagents efficiently. It is attained excluding the non-useful electrophoresis zones. For a better optimization, TAFE chambers have several useful electrophoresis zones (UEZ), each carrying a minigel.

Abstract: A process for rapid typing of yeast, parasites and bacteria is provided. It comprises the following steps: a) Preparing immobilized intact DNA in 5 to 60 minutes aided by a method involving a reagent kit that only contains buffer solution, a detergent, a metal chelating agent and an agent to disrupt the hydrogen bonds. b) Separating intact DNA molecules or their restriction fragments in miniequipments for Pulsed Field Gel Electrophoresis of the CHEF (Contour Clamped Homogeneous Electric Field) and TAFE (Transversal Alternating Field Electrophoresis) systems in times comprised between 2.5 and 7 hours. c) Selecting the optimal conditions that should be set in miniCHEF with the aid of a method to simulate the electrophoresis patterns d) Providing reorientation times, migration velocities and sizes of the molecules calculated with the aid of a method to analyze the migrated distances without the use of size markers.

Abstract: Circuit to clamp the voltages in the electrodes of the CHEF system chambers of Pulsed Field Gel Electrophoresis (PFGE) which is formed by two identical clamping circuits that are connected to a power supply through an alternator and only one of the two circuits receives electric power at the same time. Each clamping circuit is formed by several resistors and diodes connected in series to form a voltage divider. Voltage repeaters are connected to the nodes formed in the union of two resistors. Each repeater is connected to a pair of electrodes that should be polarized at the same potential. Diodes are introduced to correct small errors in the voltage pattern applied to the electrodes. The circuit is able to maintain the potential of each electrode in front of the variations of conductivity that occur inside the chamber during the electrophoresis. This way each imposition circuit generates a homogeneous electric field of same value and different direction in a PFGE chamber of the CHEF system.

Abstract: Methods of use, accessories and chambers, optimal for performing Pulsed Field Gel Electrophoresis (PFGE) of DNA molecules in ‘Contour Clamped Homogeneous Electric Field’ (CHEF) and ‘Transversal Alternating Field Electrophoresis’ (TAFE) systems, are provided herein. DNA molecules are rapidly separated in the minigels of these chambers. The sizes of chambers and accessories are determined by the separation between the opposite polarity electrodes; which is comprised between 6.2 and 15 cm. Reproducibility of molecule separation is achieved because the accessories warrant homogeneous electric resistance in the buffer and minigels. Chambers allow a high-throughput sample format using the reagents efficiently. It is attained excluding the non-useful electrophoresis zones. For a better optimization, TAFE chambers have several useful electrophoresis zones (UEZ), each carrying a minigel.

Abstract: Methods of use, accessories and chambers, optimal for performing Pulsed Field Gel Electrophoresis (PFGE) of DNA molecules in ‘Contour Clamped Homogeneous Electric Field’ (CHEF) and ‘Transversal Alternating Field Electrophoresis’ (TAFE) systems, are provided herein. DNA molecules are rapidly separated in the minigels of these chambers. The sizes of chambers and accessories are determined by the separation between the opposite polarity electrodes; which is comprised between 6.2 and 15 cm. Reproducibility of molecule separation is achieved because the accessories warrant homogeneous electric resistance in the buffer and minigels. Chambers allow a high-throughput sample format using the reagents efficiently. It is attained excluding the non-useful electrophoresis zones For a better optimization, TAFE chambers have several useful electrophoresis zones (UEZ), each carrying a minigel.

Abstract: Circuit to clamp the voltages in the electrodes of the CHEF system chambers of Pulsed Field Gel Electrophoresis (PFGE) which is formed by two identical clamping circuits that are connected to a power supply through an alternator and only one of the two circuits receives electric power at the same time. Each clamping circuit is formed by several resistors and diodes connected in series to form a voltage divider. Voltage repeaters are connected to the nodes formed in the union of two resistors. Each repeater is connected to a pair of electrodes that should be polarized at the same potential. Diodes are introduced to correct small errors in the voltage pattern applied to the electrodes. The circuit is able to maintain the potential of each electrode in front of the variations of conductivity that occur inside the chamber during the electrophoresis. This way each imposition circuit generates a homogeneous electric field of same value and different direction in a PFGE chamber of the CHEF system.

Abstract: A process for rapid typing of yeast, parasites and bacteria is provided. It comprises the following steps: a) Preparing immobilized intact DNA in 5 to 60 minutes aided by a method involving a reagent kit that only contains buffer solution, a detergent, a metal chelating agent and an agent to disrupt the hydrogen bonds. b) Separating intact DNA molecules or their restriction fragments in miniequipments for Pulsed Field Gel Electrophoresis of the CHEF (Contour Clamped Homogeneous Electric Field) and TAFE (Transversal Alternating Field Electrophoresis) systems in times comprised between 2.5 and 7 hours. c) Selecting the optimal conditions that should be set in miniCHEF with the aid of a method to simulate the electrophoresis patterns d) Providing reorientation times, migration velocities and sizes of the molecules calculated with the aid of a method to analyze the migrated distances without the use of size markers.