Introduction
Blood samples contain rich DNA, including mitochondrial DNA, genomic DNA, circulating DNA (mostly released into blood after tumor cell apoptosis) in white blood cells, as well as parasitic viral or microbial DNA. These DNA are important parameters in clinical testing or diagnosis, which are also valuable materials for medical research. There are three main issues with extracting DNA from blood samples:
1. The sample is highly infectious, posing great harm to operators and the environment.
2. The source of DNA is complex and aportion of the nucleic acid, such as viral DNA or free DNA, may be lost during the operation, leading to downstream detection failure;
3. Blood sample contains a large amount of impurities and inhibitory factors.
Currently there are many methods available for extracting DNA from whole blood samples, such as phenol chloroform extraction, salting out method, etc. However, these methods require pre-treatment of blood sample, which removes red blood cells and isolate white blood cells in the first step. Due to the requirement that it cannot inactivate or kill pathogens during the process of removing red blood cells, the waste liquid (red blood cell lysate) and consumables may be contaminated by pathogens and become infectious, posing a danger to the entire laboratory environment and operators. In addition, during the process of removing red blood cells, useful nucleic acid information such as viruses, microorganisms, or circulating DNA is also lost, leading to experiment or detection failures.
The HiPure Blood DNA Kits series provided by Magen Company uses silica gel column purification technology, which can directly lyse whole blood samples without the need for white blood cell separation. Whole blood samples are directly mixed with lysates and proteases, resulting in the inactivation of pathogens, greatly reducing the infectivity, environmental pollution, and the chance of operators being infected. Due to the direct lysis and digestion of samples, except lymphocyte DNA, other circulating DNA as well as DNA from viruses and microorganisms, can also be recovered.
This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern Blotting. Total DNA (e.g., genomic, viral, mitochondrial) can be purified from tissue and culture cells.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total DNA from 2ml blood and 200mg tissue using Midi column |
| Applications | PCR, southern bolt and virus detection, etc |
| Purification method | Midi spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Tissue, cell, blood, saliva, swab, blood spot, semen and other clinical samples |
| Sample amount | 0.2-2 ml |
| Elution volume | ≥300μl |
| Time per run | ≤80 minutes |
| Liquid carrying volume per column | 4ml |
| Binding yield of column | 1mg |
Principles
This product is based on silica column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid 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 quality DNA – meet a variety of downstream applications, including PCR, qPCR, enzyme digestion, hybridization, etc.
- Fast – without separation of leukocytes, organic extraction or ethanol precipitation
- Simple – all nucleic acids can be obtained by direct digestion
- Wide applicability – handle a variety of liquid samples
Kit Contents
| Contents | D311302 | D311303 |
| Purification Times | 20 | 100 |
| HiPure gDNA Midi Columns | 20 | 100 |
| 15ml Collection Tubes | 40 | 200 |
| Buffer ATL | 50 ml | 250 ml |
| Buffer AL | 50 ml | 250 ml |
| Buffer GW1* | 22 ml | 110 ml |
| Buffer GW2* | 12 ml | 50 ml |
| RNase A | 20 mg | 90 mg |
| Proteinase K | 100 mg | 440 mg |
| Protease Dissolve Buffer | 10 ml | 30 ml |
| Buffer AE | 20 ml | 120 ml |
Storage and Stability
Proteinase K, RNase A 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 at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
Purchase Guide
| Name | CAT NO | Sample amount | Leukocyte protocol* | Colum type | Elutio volume | Average yield | Time per run |
| HiPure Blood DNA Mini Kit | D3111 | 10-200μl | 1ml | 2ml column | ≥20μl | 5-9μg/200μl | ≤30 minutes |
| HiPure Tissue&Blood DNA Midi Kit | D3113 | 0.2-2ml | 10ml | 1.5ml column | ≥300μl | 20-40μg/1m | ≤80 minutes |
| HiPure Tissue&Blood DNA Maxi Kit | D3115 | 3 -10ml | 10ml | 15ml column | ≥700μl | 20-40μg/1m | ≤90 minutes |
| HiPure Tissue&Blood DNA 96 Kit | D3117 | 1-200μl | 1ml | 96 well plate | 3-8μg/200μl |
