Introduction
Hipure Stool RNA Kit is specially designed for stool RNA extraction. This kit is suitable for extracting high-purity microbial or host cell RNA from ≤0.1g stool samples. The kit adopts silica gel column purification technology and original solution system, which can effectively remove humic acid and other inhibitory factors in stool samples. The purified RNA can be directly used in RT-PCR, Northern hybridization and other experiments.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from 100-150mg stool sample |
| Applications | RT-PCR, Northern hybridization and other experiments |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Stool |
| Sample amount | 100-150 mg |
| Elution volume | ≥30μl |
| Time per run | ≤50 minutes |
| Liquid carrying volume per column | 100µg |
| Binding yield of column | 800µl |
Principle
The HiPure silica gel column uses a high binding ability glass fiber filter membrane as the substrate. Under the condition of high concentration of ionizing agent (such as Guanidinium chloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bonding and electrostatic and other physical factors, while protein or other impurities are not adsorbed and removed. The filter membrane that has adsorbed nucleic acids is washed to remove proteins and salts. Finally, low salt buffer solution (such as Buffer TE) or water can be used to wash out the nucleic acids adsorbed on the filter membrane. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.
The stool samples are homogenized in the lysis solution, further lysed in a high-temperature water bath, and RNA is released into the lysis solution. Chloroform extraction removes genomic DNA and impurities, transfer the supernatant to an alcohol free binding solution, purify RNA through a column, and finally elute RNA with RNase Free Water. The purified RNA can be directly used for experiments such as PCR, Southern hybridization, and enzyme digestion.
Advantages
- High purity – unique adsorbent for more efficient removal of inhibitors
- High concentration – maximum extraction of total RNA from stool samples
- Sensitive – RNA can be purified at the level of PG
Kit Contents
| Contents | R418502 | R418503 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Mini Columns | 50 | 250 |
| 2ml Collection Tubes | 50 | 250 |
| Glass Beads (0.1~0.6mm) | 30 g | 150 g |
| Buffer SPL | 30 ml | 140 ml |
| Buffer PHC | 30 ml | 140 ml |
| Buffer GRP | 60 ml | 250 ml |
| Buffer RW1 | 50 ml | 250 ml |
| Buffer RW2 * | 20 ml | 2 x 50 ml |
| RNase Free Water | 15 ml | 30 ml |
Storage and Stability
The kit components can be stored at room temperature (15–25°C) and are stable for 18 months under these conditions. At low temperatures, Buffer SPL may form precipitates, dissolve it by 55°C water bath. After receiving the product, Buffer PHC should be stored at 2-8°C.
