Introduction
This product offers a fast and reliable method to isolate RNA, including miRNA, from a wide range of samples:
- Tissue
- Cells
- Blood
- Other clinical samples
The extracted RNA is suitable for various downstream applications, including:
- RT-PCR (Reverse Transcription PCR)
- Quantitative RT-PCR (qRT-PCR)
- Viral RNA detection
- Other RNA-based analyses
This highlights the product’s versatility and compatibility with various research needs in gene expression analysis and RNA detection.
Specifications
| Features | Specifications |
| Main Functions | Isolation of total RNA（miRNA）from tissue, and cell using two columns and DNase plus reagent |
| Applications | RT-PCR, cDNA synthesis, second-generation sequencing |
| Products | RNA, miRNA |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Clinical tissues, cells, lymphocytes |
| Sample amount | Tissue: <20 mg Cells: <5 x 106 |
| Yield | 2-50μg |
| Elution volume | ≥30μl |
| Time per run | ≤25 minutes |
Principle
- Purification Method: Silica column purification is employed in this product for effective nucleic acid extraction.
- Paraffin Removal: Buffer DPS is utilized to remove paraffin from the samples prior to the purification process.
- Sample Lysis and Digestion: The sample lysis process, coupled with proteinase K digestion, takes only 15 minutes, ensuring rapid processing.
- Incubation Conditions: Following lysis, samples are subjected to a 15-minute incubation at 80°C to facilitate optimal nucleic acid release.
- Nucleic Acid Adsorption: Transfer of the lysed samples to an adsorption column allows RNA to adsorb onto the membrane selectively, while proteins remain unadsorbed and are removed through filtration.
- Washing Step: Proteins and other impurities are washed away from the membrane to enhance the purity of the extracted RNA.
- Elution: Finally, RNA is eluted from the membrane using a low-salt buffer, ensuring efficient recovery of high-quality RNA.
Advantages
- Efficient DNA removal – One-step RNA extraction can effectively remove genomic DNA
- High quality – one-step RNA extraction reagent combined with silica gel column can obtain the highest concentration
- Fast – the whole extraction only takes 15-25 minutes
- Nontoxic – no toxic phenol-chloroform extraction is required in the extraction
Kit Contents
| Contents | IVD4121 |
| Purification Times | 50 Preps |
| HiPure DNA Mini Column Ⅱ | 50 |
| HiPure RNA Mini Columns | 50 |
| 2ml Collection Tubes | 150 |
| Proteinase K | 24 mg |
| Protease Dissolve Buffer | 1.8 ml |
| DNase I | 600 μl |
| DNase Buffer | 6 ml |
| Buffer RTL | 40 ml |
| RNA Digestion Buffer | 15 ml |
| Buffer RWC* | 20 ml |
| Buffer RW2* | 20 ml |
| Nuclease Free Water | 10 ml |
Storage and Stability
- Proteinase K Storage: Store Proteinase K at 2–8°C upon arrival. Short-term storage (up to 8 weeks) at room temperature (15–25°C) is acceptable without affecting its performance.
- DNase I Storage: Store DNase I at -20°C. Short-term storage (up to 1 week) at room temperature (15–25°C) does not affect its performance.
- Remaining Kit Components Storage: The remaining kit components can be stored at room temperature (15–25°C) and remain stable for at least 18 months under these conditions.
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