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).
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 |
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 |
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:
Mastitis is the single most costly disease of dairy cattle resulting in the reduction of milk yield and quality. The inflammation of the utter is mainly caused by infection of various bacteria. Streptococcus uberis is a gram-positive bacterium that is known worldwide as an environmental pathogen responsible for a high proportion of cases of mastitis in lactating cows and is also the predominant organism isolated from mammary glands during the non-lactating period. Often it is resistant to treatment and causes persistent high somatic cell counts without clinical mastitis.
Streptococcus uberis TaqMan PCR Kit, 100 reactions
Streptococcus uberis TaqMan PCR Probe/Primer Set and Controls, 100 reactions
For research use only and NOT intended for in vitro diagnostics.
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Storage Conditions and Product Stability
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
| Component | Cat. TM30850 (100 preps) | Cat. TM30810 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| S. uberis Primer & Probe Mix | 280 μL | 280 μL |
| S. uberis Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
Bile Salt Export Pump (BSEP) is a member of the ATP-binding cassette (ABC) transporters, which mediates the transport of bile acid, taurocholate and other cholate conjugates across the hepatocyte canalicular membrane into the canaliculus. BSEP is associated with progressive familial intrahepatic cholestasis type 2 (PFIC2) and benign recurrent intrahepatic cholestasis type 2 (BRIC2). PFIC2 caused by mutations in the BSEP gene increases the risk of hepatocellular carcinoma in early life.
