Introducción
- Rápido, Simple, and Cost-Effective: Offers a rapid, easy-to-use, and economical method for ADN plásmido miniprep suitable for routine molecular biology applications.
- Silica Membrane Technology: Utilizes silica membrane technology, eliminating the need for cumbersome steps associated with loose resins or slurries, streamlining the purification process.
- Immediate Usability: Purified plasmid DNA is immediately ready for downstream applications, ahorrando tiempo y esfuerzo.
- No Phenol Extraction or Ethanol Precipitation: Bypasses the need for phenol extraction and ethanol precipitation, simplifying the workflow and reducing hands-on time.
- ADN de alta calidad: Yields high-quality plasmid DNA eluted in a small volume of Tris buffer or water, ensuring optimal performance in downstream applications.
Especificaciones
| Características | Especificaciones |
| Funciones principales | Isolation of up to 35μg plasmid DNA from 1-5ml bacterial culture |
| Aplicaciones | digestión enzimática, secuenciación, PCR, cloning, etc.. |
| Método de purificación | Mini columna de giro |
| Tecnología de purificación | tecnología de sílice |
| método de proceso | Manual (centrifugación o vacío) |
| Tipo de ejemplo | Plásmido convencional, plásmido de menos de 30 KB |
| Cantidad de muestra | High copy plasmid: 1-5ml culture medium Low copy number plasmid: 5-10ml culture medium |
| Producir | 5-35µg |
| Volumen de elución | ≥30μl |
| Tiempo por carrera | Complete 1-24 samples in 30 minutos |
| Volumen de transporte de líquido por columna | 800l |
| Rendimiento de unión de la columna | 35µg |
Principio
- Alkaline Lysis: Begins with the alkaline lysis of bacterial cells, which breaks down the cell membrane and releases plasmid DNA into the lysate.
- Silica-Based DNA Binding: The released DNA is adsorbed onto the silica membrane in the presence of high salt, facilitating efficient binding of DNA to the membrane surface.
- Three Basic Steps: The procedure involves three main steps: (1) Preparation and clearing of the bacterial lysate using an alkaline method, (2) Transfer of the cleared lysate to a column for DNA binding, y (3) Washing to remove proteins and other impurities.
- Silica Membrane: Utilizes a unique silica membrane in the kit, which completely replaces traditional glass or silica slurries typically used in plasmid DNA minipreps.
- Elution with Low-Salt Buffer: After washing to remove impurities, ácido nucleico, including plasmid DNA, is eluted from the silica membrane using a low-salt buffer containing 10mM Tris, pH 9.0, and 0.5mM EDTA.
Ventajas
- Alta pureza – El plásmido purificado se puede utilizar directamente en la secuenciación., enzyme digestion PCR, etc..
- Rápido – solo se necesita 1 minute to obtain supernatant by optimized solution (10 minutes for other brands)
- Alto rendimiento – up to 35µg plasmid can be bound in one column
Contenido del kit
| Contenido | P100102 | P100103 |
| Tiempos de purificación | 100 preparativos | 250 preparativos |
| ARNasa A | 5 mg | 10 mg |
| Búfer P1 | 30 ml | 80 ml |
| Tampón P2 | 30 ml | 80 ml |
| Buffer P3 | 40 ml | 100 ml |
| Búfer PW1 | 60 ml | 140 ml |
| Búfer PW2* | 20 ml | 50 ml |
| Tampón de elución | 15 ml | 30 ml |
| Minicolumnas HiPure DNA II | 100 | 250 |
| 2 Tubos de recolección de ml | 100 | 250 |
Almacenamiento y estabilidad
- Condiciones de almacenaje: Kit components can be stored dry at room temperature (15-25°C) and remain stable for a minimum of 18 meses en estas condiciones.
- Buffer Precipitates: In case of any precipitates forming in the buffers, warm them at 37°C to dissolve effectively.
- Buffer P1 Stability: After the addition of RNase A, Buffer P1 remains stable for up to 6 meses cuando se almacena a 2-8°C.
Datos del experimento
Guía de compra
| Nombre | NO GATO | Cantidad de muestra | Tipo de columna | Volumen de elución | Producir | Tiempo por carrera | Requisitos de centrífuga |
| Mini kit de plásmido HiPure | P1001 | 1-5ml | 1.5columna de ml | 30-50µl | 5–35 µg/5 ml | ≤30 minutos | pequeña centrífuga |
| Mini kit II de plásmido HiPure | P1002 | 5-15ml | 1.5columna de ml | 75-100µl | 20-80µg/15ml | ≤30 minutos | pequeña centrífuga |
| HiPure Plásmido Plus 96 Equipo | P1006 | 1-1.5ml | 96 buen plato | 70-150µl | 1-15µg/1ml | ≤60 minutos | 96-centrífuga de placa de pocillos |
| Kit Midi de plásmido HiPure Fastfilter | P1013 | 30-50ml | 15columna de ml | 0.3-0.8ml | 50–250 µg/50 ml | ≤60 minutos | 15centrífuga ml |
| Kit maxi de plásmido HiPure Fastfilter | P1014 | 100-200ml | 50columna de ml | 0.5-1.5ml | 0.4–1mg/200ml | ≤60 minutos | 50centrífuga ml |
Reseñas
Aún no hay reseñas.