Introduction
- Purpose: Extraction of stool RNA.
- Sample Compatibility: Suitable for extracting high-purity microbial or host cell RNA from ≤0.1g stool samples.
- Technology: Utilizes silica gel column purification technology and an original solution system.
- Key Feature: Effectively removes humic acid and other inhibitory factors present in stool samples.
- Applications: The purified RNA can be directly used in RT-PCR, Northern hybridization, and other experiments.
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
- Principle of HiPure Silica Gel Column: Utilizes a high binding ability glass fiber filter membrane as the substrate. Under high concentrations of ionizing agents (e.g., Guanidinium chloride or guanidine isothiocyanate), nucleic acids are adsorbed through hydrogen bonding and electrostatic interactions, while proteins and other impurities are not adsorbed and removed. Subsequently, the filter membrane with adsorbed nucleic acids is washed to remove proteins and salts. Finally, low salt buffer solution (e.g., Buffer TE) or water is used to elute the purified nucleic acids, resulting in high purity suitable for downstream experiments.
- Sample Processing: Stool samples are homogenized in the lysis solution, then further lysed in a high-temperature water bath to release RNA into the solution. Chloroform extraction is employed to remove genomic DNA and impurities. The supernatant is transferred to an alcohol-free binding solution, followed by RNA purification through a column. Finally, RNA is eluted using RNase Free Water.
- Applications: The purified RNA is suitable for various downstream experiments such as PCR, Southern hybridization, and enzyme digestion.
Advantages
- High Purity: Utilizes a unique adsorbent for more efficient removal of inhibitors, ensuring high purity of extracted RNA.
- High Concentration: Achieves maximum extraction of total RNA from stool samples due to its high concentration, maximizing RNA yield.
- Sensitive: Capable of purifying RNA at the level of picograms (PG), ensuring sensitivity in downstream applications.
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
- Storage Conditions: Kit components can be stored at room temperature (15–25°C) and remain stable for 18 months under these conditions.
- Buffer SPL: May form precipitates at low temperatures. Dissolve precipitates by placing the Buffer SPL in a 55°C water bath.
- Buffer PHC: After receiving the product, store Buffer PHC at 2-8°C.
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