Introduction
Hipure Microbial RNA kit is suitable for extracting high-purity total RNA from bacterial/yeast culture medium. This kit combines high-efficiency Magzol Reagent (one-step extraction reagent) and silicagel column purification technology to complete the extraction of high-purity total RNA in only 40 minutes. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from bacteria, yeast cells |
| Applications | RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Bacteria, yeast cells |
| Sample amount | Bacteria: <10^9；Yeast cells：<2 x10^7 |
| Elution volume | ≥30μl |
| Time per run | ≤40 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
Principle
Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of a high concentration of ionizing agent (such as guanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein and other impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.
Advantages
- Fast – several samples can be extracted in 30 minutes
- High purity – the purified RNA can be directly used in various downstream applications
- High recovery – RNA can be recovered at the level of PG
- Good repeatability – silica gel column purification technology can obtain ideal results every time
- Broad spectrum – it can deal with various bacteria, including Gram-positive bacteria that are difficult to be lysed
- Sufficient components – the kit contains carried wall-breaking enzymes and glass beads
Kit Contents
| Contents | R418202 | R418203 |
| 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 |
| DNase I | 600 μl | 5 x 600 μl |
| DNase Buffer | 6 ml | 30 ml |
| Protease Dissolve Buffer | 1.8 ml | 15 ml |
| Buffer ATL | 50 ml | 200 ml |
| Buffer PHC | 50 ml | 200 ml |
| Buffer GXP2* | 20 ml | 100 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
Buffer PHC should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Buffer PHC up to 12 weeks) at room temperature(15–25°C) does not affect their performance. The remaining kit components can be stored at roomtemperature (15–25°C) and are stable for at least 18 months under these conditions.
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