HiPure DNA Clean Up Kit usesproprietary chemistry and HiPure technology to recover DNA Fragments between20bp-20kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing,restriction digestion, or various labeling reactions. In addition, this kit canbe also used to recover DNA directly from enzymatic reactions such as PCR andenzyme digestion reactions.
Specifications
| Features | Specifications |
| Main Functions | Recover DNA fragments between 20bp-20kbp from crude DNA and enzymatic reaction solution |
| Applications | PCR, sequencing, labeling reactions, ligations and restriction digestion, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Crude DNA, enzymatic reaction solution |
| Sample amount | <150μl |
| Recovery | 80% |
| Elution volume | ≥15μl |
| Time per run | ≤10 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 20µg |
The HiPure system uses a simple bind-wash-elute procedure. Gelslices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding, and the mixture is applied to the column. Nucleic acids adsorb to the silica-gel membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure DNA is eluted with a small volume of low-salt buffer provided or water, ready to use in subsequent applications.
Advantages
Kit Contents
| Contents | D214102 | D214103 |
| Purification Times | 100 Preps | 250 Preps |
| Buffer GWP | 30 ml | 60 ml |
| Buffer DW2 | 20 ml | 50 ml |
| Elution Buffer | 30 ml | 60 ml |
| HiPure DNA Mini Columns I | 50 | 250 |
| 2 ml Collection Tubes | 50 | 250 |
Storage and Stability
HiPure DNA Clean Up Kit components are guaranteed for at least one year when stored at room temperature. If any precipitates form in the buffers, warm at 37℃ to dissolve.
HiPure DNA Clean Up Kit usesproprietary chemistry and HiPure technology to recover DNA Fragments between20bp-20kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing,restriction digestion, or various labeling reactions. In addition, this kit canbe also used to recover DNA directly from enzymatic reactions such as PCR andenzyme digestion reactions.
These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used, as heparin can significantly interfere with many downstream applications such as RT-PCR. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS.
Cell-free circulating RNA, including exosomal RNA in plasma or serum, has the potential to provide biomarkers for certain cancers and disease states, and includes tumor-specific extracellular RNA in the blood. Exosomes are 40 – 100 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which depend upon the tumour cell type from which they are secreted. For this reason, exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses, they can be efficiently recovered from biological fluids, such as plasma or serum.
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 μL to 200 μL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 μL to 25 μL.
This utilizes a two-column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 μL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
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| Kit Specifications | |
| Sample Type | Plasma/Serum |
| Anti-Coagulant (for Plasma)† | EDTA or Citrate |
| Sample Volume Range | 50 to 200 μL |
| Minimum Elution Volume | 10 μL |
| Maximum Elution Volume | 25 μL |
| Time to Complete 10 Purifications | 15 – 20 minutes |
| Size of RNA Purified | All sizes, including miRNA and small RNA (<200 nt) |
| Average Yields¥ | Variable depending on specimen |
† This kit is suitable for the isolation of RNA from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR
¥ Please check page 5 for Average Plasma/Serum Yields and Common RNA Quantification Methods
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
| Component | Cat. 73700 (50 preps) | Cat. 73710 (20 preps) | Cat. 73720 (10 preps) |
|---|---|---|---|
| Lysis Buffer A | 2 x 20 mL | 100 mL | 1 x 130 mL 1 x 30 mL |
| Wash Solution A | 18 mL* | 1 x 38 mL* 1 x 18 mL* | 38 mL* |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Elution Buffer F | – | – | 15 mL |
| EXTRACLean Micro Spin Columns | 50 | – | – |
| EXTRACLean Mini Spin Columns | – | 20 | 10 |
| EXTRACLean Midi Spin Columns | – | 20 | – |
| EXTRACLean Maxi Spin Columns | – | – | 10 |
| Collection Tubes | – | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 |
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 whole blood, tissue and culture cells.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total DNA from 10ml blood and 1g tissue using Maxi column |
| Applications | PCR, southern bolt and virus detection, etc |
| Purification method | Maxi 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 | 3-10ml |
| Elution volume | ≥700μl |
| Time per run | ≤90 minutes |
| Liquid carrying volume per column | 4ml |
| Binding yield of column | 5mg |
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 | D311502 | D311503 |
| Purification Times | 10 | 50 |
| HiPure gDNA Maxi Columns | 10 | 50 |
| 50ml Collection Tubes | 20 | 100 |
| Buffer ATL | 120 ml | 550 ml |
| Buffer AL | 120 ml | 2 x 300 ml |
| Buffer GW1* | 53 ml | 220 ml |
| Buffer GW2* | 25 ml | 110 ml |
| RNase A | 40 mg | 180 ml |
| Proteinase K | 120 mg | 540 mg |
| Protease Dissolve Buffer | 12 ml | 50 ml |
| Buffer AE | 30 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.
| 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 |
*Note：Leukocyte protocol can be used when large volume whole blood samples need to be processed. Whole blood was treated with red blood cell lysate, and white blood cells were obtained by centrifugation before extraction
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: