Introducción
Hipure viral RNA kit is suitable for purifying viral RNA from samples such as cell-free body fluid or culture medium. The kit is based on silica gel column purification technology. It requires no toxic phenol chloroform extraction and time-consuming alcohol precipitation in the extraction. The whole extraction process takes only 25 minutos. The kit is suitable for extracting viral RNA from 1-140µl cell-free fluid samples such as serum, plasma, cell-free body fluid or culture medium. The product has successfully extracted hepatitis B A/C, hepatitis C RNA, SARS and HIV. The obtained RNA can be directly used for RT-PCR, Northern hybridization and virus detection.
Detalles
Especificaciones
| Características | Especificaciones |
| Funciones principales | Extract viral RNA from 140μl cell-free samples |
| Aplicaciones | RT-PCR, hibridación del norte, and various virus detection |
| Método de purificación | Mini columna de giro |
| Tecnología de purificación | tecnología de sílice |
| método de proceso | Manual (centrifugación o vacío) |
| Tipo de ejemplo | Cell-free body fluid or culture medium |
| Cantidad de muestra | 140µl |
| Volumen de elución | ≥15μ |
| Tiempo por carrera | ≤25 minutos |
| Volumen de transporte de líquido por columna | 800µl |
| Rendimiento de unión de la columna | 100µg |
Principio
Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (como guanidina hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein and other impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.
The sample is homogenized and lysed in the lysate, and RNA is released into the lysate. The high concentration of guanidine isothiocyanate contained in the lysate denatured and inactivated endogenous or exogenous RNase, while RNA is protected from degradation. The lysate is centrifuged to remove insoluble impurities. After adding ethanol to adjust the binding conditions, it is transferred to the column for filtration. RNA is adsorbed on the membrane of the column, while protein is removed without adsorption. The column is washed with buffer VHB to remove protein and other impurities, washed with buffer RW2 to remove salt. Finally the RNA is eluted by RNase free water. The eluted RNA can be directly used in RT-PCR, Northern blot, poly-A purification, traducción in vitro, etc..
Ventajas
- Rápido – Se pueden extraer varias muestras en 20 minutos por método de columna
- Alta calidad – El ARN total de alta pureza se puede utilizar directamente en diversas aplicaciones sensibles posteriores.
- Seguro – no se requiere extracción con fenol cloroformo
- Sensible – El ARN se puede recuperar a nivel de PG
- Alto rendimiento – carrier RNA contained in the product maximize the recovery of trace nucleic acid
Contenido del kit
| Contenido | R417102 | R417103 |
| Tiempos de purificación | 50 preparativos | 250 preparativos |
| HiPure Viral Micro Columns | 50 | 250 |
| 2Tubos de recolección de ml | 100 | 500 |
| Buffer VRL | 50 ml | 200 ml |
| Carrier RNA | 310 µg | 3 X 310 µg |
| Buffer VHB* | 13 ml | 110 ml |
| Búfer RW2* | 20 ml | 2 X 50 ml |
| Agua libre de nucleasas | 10 ml | 30 ml |
Almacenamiento y estabilidad
Carrier RNA should be stored at 2–8°C upon arrival. Sin embargo, almacenamiento a corto plazo (hasta 12 semanas) a temperatura ambiente (15–25°C) no afecta su rendimiento. Los componentes restantes del kit se pueden almacenar a temperatura ambiente. (15–25°C) y son estables durante al menos 18 months under theseconditions
Reseñas
Aún no hay reseñas.