Introduction
The HiPure Liquid RNA & miRNA Kit integrates phenol/guanidine-based sample lysis and silica membrane purification of total RNA. MagZol LS Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate designed to facilitate lysis of liquid sample and inhibit RNase. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable the use of up to 0.25ml liquid sample per mini spin column.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from 0.25ml body fluids using column and MagZol LS reagent |
| 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 | Blood, serum, plasma |
| Sample amount | 250μl |
| Elution volume | ≥20μl |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100μg |
Principle
0.25ml liquid samples are homogenized in 0.75ml MagZol LS Reagent. After addition of chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. The upper, aqueous phase is extracted, and ethanol is added to provide appropriate binding conditions. The sample is then applied to the spin column, where the total RNA (up to 100 µg) binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in 30–100µl of RNase-free water.
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 40 minutes by using column purification method
- Safety – no phenol chloroform extraction required
- Sensitive – direct lysis of blood, plasma and other samples without separation of leukocytes
Kit Contents
| Contents | R416302 | R416303 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Mini Columns | 50 | 250 |
| 2ml Collection Tubes | 50 | 2 x 125 |
| MagZol LS Reagent | 60 ml | 270 ml |
| Buffer RWC | 20 ml | 60 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 30 ml |
Storage and Stability
MagZol LS Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
