CU22849A1 - CHAMBER OF ELECTROPHOROSIS OF PULSING FIELDS, ACCESSORIES AND METHODS OF USE FOR THE QUICK AND REPRODUCIBLE SEPARATION OF LARGE DNA MOLECULES - Google Patents
CHAMBER OF ELECTROPHOROSIS OF PULSING FIELDS, ACCESSORIES AND METHODS OF USE FOR THE QUICK AND REPRODUCIBLE SEPARATION OF LARGE DNA MOLECULESInfo
- Publication number
- CU22849A1 CU22849A1 CU20000306A CU20000306A CU22849A1 CU 22849 A1 CU22849 A1 CU 22849A1 CU 20000306 A CU20000306 A CU 20000306A CU 20000306 A CU20000306 A CU 20000306A CU 22849 A1 CU22849 A1 CU 22849A1
- Authority
- CU
- Cuba
- Prior art keywords
- electrophoresis
- chambers
- dna molecules
- mini
- field
- Prior art date
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
La presente invención se relaciona con el campo de la Biología Molecular y en particular se refiere a mini-cámaras de electroforesis de campos pulsantes de los sistemas `Contour Clamped Homogeneous Electric Field' o `Transversal Alternating Field Electrophoresis' (TAFE) y su método de empleo. Las cámaras de la presente invención se emplean para la separación de grandes moléculas de ADN mediante electroforesis de campos pulsantes (ECP) en mini equipos y geles. Las mini-cámaras de esta invención poseen un sistema para limitar la formación de turbulencias en la solución tampón, para preparar geles planos, para preparar los bloques que contienen las muestras y para alinear las muestras. Estos sistemas garantizan que el área de sección tranversal al paso de la corriente, tanto por el gel como por la solución tampón donde se sumerge el gel, sea constante en toda la cámara de electroforesis e impida modificaciones regionales de la corriente que fluye en la misma, de aquí que la migración de las moléculas de ADN de cualquier talla no se modifica incontroladamente durante el proceso de electroforesis. Las cámaras además poseen un sistema para fijar y mantener tensos los electrodos. De esta forma se simplifica su operación y se contribuye a la reproducibilidad de los patrones de bandas. Con estas cámaras se obtienen separaciones reproducibles de moléculas de hasta 2 Mb en menos de 5 horas y de hasta 1 Mb en menos de 2.5 horas cuando se emplean campos eléctricos entre 8-16 V/cm, 20C, así como la duración adecuada de los pulsos eléctricos y una correcta polarización de los electrodos. En la invención se brinda además un método para seleccionar las condiciones de electroforesis empleando dichas cámaras.The present invention relates to the field of Molecular Biology and in particular relates to pulsed field electrophoresis mini-chambers of the 'Contour Clamped Homogeneous Electric Field' or 'Transversal Alternating Field Electrophoresis' (TAFE) systems and their method of job. The chambers of the present invention are used for the separation of large DNA molecules by pulsed field electrophoresis (ECP) in mini equipment and gels. The mini-chambers of this invention have a system to limit the formation of turbulence in the buffer solution, to prepare flat gels, to prepare the blocks containing the samples and to align the samples. These systems ensure that the cross-sectional area to the passage of the current, both by the gel and by the buffer solution where the gel is submerged, is constant throughout the electrophoresis chamber and prevents regional modifications of the current flowing in it , hence the migration of DNA molecules of any size does not change uncontrollably during the electrophoresis process. The cameras also have a system to fix and keep the electrodes tight. This simplifies its operation and contributes to the reproducibility of band patterns. With these cameras reproducible separations of molecules of up to 2 Mb in less than 5 hours and up to 1 Mb in less than 2.5 hours are obtained when electric fields are used between 8-16 V / cm, 20C, as well as the adequate duration of the electrical pulses and correct polarization of the electrodes. The invention also provides a method for selecting electrophoresis conditions using said chambers.
Priority Applications (11)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CU20000306A CU22849A1 (en) | 2000-12-27 | 2000-12-27 | CHAMBER OF ELECTROPHOROSIS OF PULSING FIELDS, ACCESSORIES AND METHODS OF USE FOR THE QUICK AND REPRODUCIBLE SEPARATION OF LARGE DNA MOLECULES |
| DE60140190T DE60140190D1 (en) | 2000-06-07 | 2001-06-07 | ELECTROPHORESE CHAMBER IN PULSATING FIELD, ACCESSORIES AND METHOD FOR USE FOR THE SEPARATION OF DNA MOLECULES |
| PT01942956T PT1291649E (en) | 2000-06-07 | 2001-06-07 | Pulsed field electrophoresis chambers, accessories and method of utilization for separation of dna molecules |
| PCT/CU2001/000003 WO2001094932A1 (en) | 2000-06-07 | 2001-06-07 | Pulsed field electrophoresis chambers, accessories and method of utilization for separation of dna molecules |
| EP01942956A EP1291649B1 (en) | 2000-06-07 | 2001-06-07 | Pulsed field electrophoresis chambers, accessories and method of utilization for separation of dna molecules |
| US10/070,878 US7189316B2 (en) | 2000-06-07 | 2001-06-07 | Pulsed field electrophoresis chambers, accessories and method of utilization for separation of DNA molecules |
| MXPA02002259A MXPA02002259A (en) | 2000-06-07 | 2001-06-07 | Pulsed field electrophoresis chambers, accessories and method of utilization for separation of dna molecules. |
| ES01942956T ES2334978T3 (en) | 2000-06-07 | 2001-06-07 | ELECTROPHORESIS CAMERAS OF PULSING FIELDS, ACCESSORIES AND EMPLOYMENT PROCEDURE FOR THE SEPARATION OF DNA MOLECULES. |
| AU65746/01A AU6574601A (en) | 2000-06-07 | 2001-06-07 | Pulsed field electrophoresis chambers, accessories and method of utilization for seperation of DNA molecules |
| AT01942956T ATE445835T1 (en) | 2000-06-07 | 2001-06-07 | ELECTROPHORESIS CHAMBER IN PULSATING FIELD, ACCESSORIES AND METHOD USED FOR SEPARATION OF DNA MOLECULES |
| US11/643,454 US8034225B2 (en) | 2000-06-07 | 2006-12-21 | Pulsed field gel electrophoresis chambers, accessories and methods of use for the separation of DNA molecules |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CU20000306A CU22849A1 (en) | 2000-12-27 | 2000-12-27 | CHAMBER OF ELECTROPHOROSIS OF PULSING FIELDS, ACCESSORIES AND METHODS OF USE FOR THE QUICK AND REPRODUCIBLE SEPARATION OF LARGE DNA MOLECULES |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CU22849A1 true CU22849A1 (en) | 2003-04-28 |
Family
ID=46599437
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CU20000306A CU22849A1 (en) | 2000-06-07 | 2000-12-27 | CHAMBER OF ELECTROPHOROSIS OF PULSING FIELDS, ACCESSORIES AND METHODS OF USE FOR THE QUICK AND REPRODUCIBLE SEPARATION OF LARGE DNA MOLECULES |
Country Status (1)
| Country | Link |
|---|---|
| CU (1) | CU22849A1 (en) |
-
2000
- 2000-12-27 CU CU20000306A patent/CU22849A1/en unknown
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20050167270A1 (en) | Electrophoresis apparatus and method | |
| Righetti et al. | Preparative purification of human monoclonal antibody isoforms in a multi-compartment electrolyser with immobiline membranes | |
| US5336387A (en) | Electrical separator apparatus and method of counterflow gradient focusing | |
| Ro et al. | Poly (dimethylsiloxane) microchip for precolumn reaction and micellar electrokinetic chromatography of biogenic amines | |
| DE60112276T2 (en) | ELECTROPHORETIC SEPARATION OF COMPOUNDS | |
| DE69109589D1 (en) | Multifunctional electrical disconnection device and separation process. | |
| JPH0781987B2 (en) | Isoelectric focusing method and apparatus therefor | |
| US3879280A (en) | Gel slab electrophoresis cell and electrophoresis apparatus utilizing same | |
| US20030019753A1 (en) | Multi-port separation apparatus and method | |
| ATE479090T1 (en) | ELECTROPHORESIS METHOD WITH PARALLEL AND SIMULTANEOUS SEPARATION | |
| US4834862A (en) | Ampholyte separation method and apparatus | |
| ATE445835T1 (en) | ELECTROPHORESIS CHAMBER IN PULSATING FIELD, ACCESSORIES AND METHOD USED FOR SEPARATION OF DNA MOLECULES | |
| Lee et al. | Characterization of electric charges in microporous membranes | |
| US4824547A (en) | Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots | |
| CU22849A1 (en) | CHAMBER OF ELECTROPHOROSIS OF PULSING FIELDS, ACCESSORIES AND METHODS OF USE FOR THE QUICK AND REPRODUCIBLE SEPARATION OF LARGE DNA MOLECULES | |
| Hirano et al. | Two‐dimensional gel electrophoresis using immobilized pH gradient tube gels | |
| Woodworth et al. | An improved vertical polyacrylamide gel electrophoresis apparatus: Application to typing and subtyping of haptoglobins | |
| US7250100B2 (en) | Two dimensional electrophoresis cassette | |
| FAWCETT et al. | Electrofocusing in wide pH range Immobiline gels containing carrier ampholytes | |
| US7025864B2 (en) | Method and apparatus for recovering target molecules from a gel containing said target molecules | |
| Lim et al. | Rapid isoelectric trapping in a micropreparative‐scale multicompartment electrolyzer | |
| JPH10500489A (en) | Method for desalting biological samples to remove interfering substances in isoelectric focusing | |
| Gianazza et al. | Fractionation of carrier ampholytes for isoelectric focusing | |
| Fawcett et al. | Simplified procedure for the preparation of immobilized pH gradient gels | |
| US6841055B2 (en) | Method of preparing electrophoretic support, electrophoretic matrix, and method of electrophoresis |