Introduction
HiPure Bacterial RNA Kit uses silica gel column purification to simplify the extraction. The whole process does not require phenol chloroform extraction and time-consuming alcohol precipitation. The kit is suitable for efficiently extracting RNA from various bacterial samples. The purified RNA can be directly used for RT-PCR, Northern hybridization, etc. The kit has included lysozyme and glass beads, which can be used to treat gram-negative bacteria which is easy to be lysed, as well as gram-positive bacteria which is hard to be lysed, including enterococcus faecalis, staphylococcus, etc.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from bacteria culture |
| 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 culture |
| Sample amount | Bacteria: <10^9 |
| 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 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 lysozyme and glass beads
Kit Contents
| Contents | R418102 | R418103 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Mini Columns | 50 | 250 |
| 2ml Collection Tubes | 50 | 250 |
| Glass Beads (0.1-0.2mm) | 20 g | 90 g |
| Lysozyme | 90 mg | 400 mg |
| Protease Dissolve Buffer | 5 ml | 15 ml |
| Buffer STE | 10 ml | 30 ml |
| MagZol Reagent | 60 ml | 270 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 30 ml |
Storage and Stability
MagZol Reagent and Lysozyme should be stored at 2-8°C upon arrival. 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. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
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