Introduction
This product is designed for purification of high-molecular-weight genomic or mitochondrial DNA from a variety of tissue and cultured cells. The convenient, scalable purification procedure removes contaminants and enzyme inhibitors such as proteins and divalent cation, and purified DNA is ready for immediate use in sensitive downstream applications or for archiving.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total DNA from tissue using economic salt out method |
| Applications | PCR, enzyme digestion, Southern hybridization,etc. |
| Purification technology | Salting out method |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Animal tissue |
| Sample amount | Unlimited |
| Elution volume | ≥100μl |
| Time per run | Variation with sample amount |
Principles
Cells are lysed with an anionic detergent in the presence of a DNA stabilizer. The DNA stabilizer limits the activity of intracellular DNases and also DNases found elsewhere in the environment. RNA is then removed by treatment with an RNA digesting enzyme. Other contaminants, such as proteins, are removed by salt precipitation. Finally, the genomic DNA is recovered by precipitation with alcohol and dissolved in Buffer TE. Purified DNA typically has an A260/A280 ratio between 1.7 and 1.9, and is up to 200 kb in size. The DNA can be safely stored at 2-8°C, -20°C, or -80°C.
Advantages
- Safe - without phenol chloroform extraction
- Environment friendly - the reagents used are safe, non-toxic and without pollution
- High molecular weight - the molecular weight of genomic DNA is about 50-150kb
- High purity - the purified DNA has A260/280=1.8-1.9, A260/230=2.0-2.5
- Unlimited sample size - solution type operation, sample volume can be adjusted at will
- Cost performance - the most economical nucleic acid extraction technology
Kit Contents
| Contents | D331201 | D331202 | D331203 |
|
Scale - grams of sample ( number of preps) |
1 g (100preps) |
5 g (500preps) |
50 g (5000preps) |
| Proteinase K | 330 µl | 1.8 ml | 18 ml |
| Buffer WTL | 33 ml | 160 ml | 2 x 800 ml |
| Buffer PPS | 12 ml | 55 ml | 500 ml |
| RNase Solution |
330 µl | 1.8 ml | 18 ml |
| Buffer TE | 12 ml | 60 ml | 200 ml |
Storage and Stability
RNase Solution should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. 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.
