Introduction
The Kit provides fast purification of high-quality RNA from whole blood, cells and tissues using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol / chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. The purified RNA can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection and in vitro translation.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from 1-1.5ml whole blood |
| Applications | qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection |
| Purification method | Mini spin column |
| Purification technology | Silica technology, acidphenol / guanidine extraction technology (MagZol pretreatment technology) |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Fresh or frozen blood, mammalian blood |
| Sample amount | ≤1.5 ml whole blood |
| Yield | 2-100μg |
| Elution volume | ≥20μl |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100μg |
Principle
The Kit simplifies isolation of RNA from blood with a fast spin-column procedure. Red blood cells are selectively lysed and white cells collected by centrifugation. White cells are then lysed using highly denaturing conditions which immediately inactivate RNases. After homogenization using the DNA spin column, the sample is applied to the RNA column. Total RNA binds to the membrane and contaminants are washed away, leaving pure RNA to be eluted in 30–100µl RNase-free water (provided with the kit) for direct use in any downstream application.
Advantages
- High quality – high purity total RNA can be directly used in various sensitive downstream applications
- Fast – isolation of several samples can be completed in 20 minutes by using column purification method
- Sensitive – RNA can be purified at the level of PG
Kit Contents
| Contents | R416102 | R416103 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure DNA Mini Columns | 50 | 250 |
| HiPure RNA Mini Columns I | 50 | 250 |
| 2ml Collection Tubes | 100 | 500 |
| 10 x Buffer RBC | 50 ml | 3 x 100 ml |
| RTL Lysis Buffer | 50 ml | 250 ml |
| Buffer RW1 | 50 ml | 250 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 30 ml |
Storage and Stability
HiPure Blood RNA Mini Kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. Make sure that all buffers are at room temperature when used. During shipment, crystals or precipitation may form in the RTL Lysis Buffer. Dissolve by warming buffer to 37°C.
