Introduction
This product provides an easy-to-use workflow for selective isolation of bacterial DNA from samples that are intrinsically rich in host DNA, such as body fluids or swabs. The method is specific for the identification of intact bacteria so it prevents false results due to nucleic acids from dead bacteria. The Kit allows isolation of enriched bacterial DNA suitable for a variety of applications, including qPCR and whole metagenome or 16S rRNA gene sequencing.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation gDNA from biological sample and remove host DNA |
| Applications | PCR, southern blot and enzyme digestion, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Culture medium, swab, parasitic blood, tissue, sputum, etc. |
| Sample amount | Blood sample, homogenate, plasma, brain effusion, swab immersion solution, centrifuged concentrated liquid, etc.:0.5-1ml |
| Elution volume | ≥50μl |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 800μl |
| Binding yield of column | 100μg |
Principle
This product is based on silica Column purification. This product efficiently depletes human and animal host DNA and yields enriched bacterial DNA. An optimized combination of mechanical and chemical lysis allows efficient disruption of bacterial cells. Target DNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
- High concentration – the host DNA is digested by dnase, without contamination of host DNA
- High recovery – DNA can be recovered at the level of PG
- Good repeatability – silica technology can obtain ideal results every time
- Wide applicability – it can be used in blood, tissue, intestinal contents and other samples
Kit Contents
| Contents | D314802 | D314803 |
| Purification Times | 50 Preps | 250 Preps |
| Hipure DNA Mini Columns I | 50 | 2 x 125 |
| 2ml Collection Tubes | 50 | 2 x 125 |
| 2ml beads Tubes | 50 | 250 |
| Buffer DRB | 15 ml | 60 ml |
| Buffer ES | 6 ml | 30 ml |
| Reagent DX | 0.5 ml | 1 ml |
| Buffer DL | 30 ml | 120 ml |
| Buffer GW1 | 22 ml | 110 ml |
| Buffer GW2 | 12 ml | 50 ml |
| DNase I (Powder) | 6 mg | 30 mg |
| Proteinase K | 24 mg | 120 mg |
| Protease Dissolve Buffer | 5 ml | 15 ml |
| Buffer AE | 15 ml | 60 ml |
Storage and Stability
DNase I and Proteinase K should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 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.
