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, whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.
Specifications
| Features | Specifications |
| Main Functions | Isolation total DNA from blood, tissue, culture cells, swab, blood spots using 96 plate |
| Applications | PCR, southern bolt and virus detection, etc |
| Purification method | 96 well plate |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Blood, serum, plasma, milk, saliva, and other liquid samples and cultured cells |
| Sample amount | |
| Elution volume | |
| Time per run | |
| Liquid carrying volume per column | |
| Binding yield of column |
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).
| Contents | D311701 | D311702 |
| Purification Times | 1 x 96 | 4 x 96 |
| HiPure gDNA Plate | 1 | 4 |
| 96 well Plate (2.2ml) | 1 | 4 |
| 1.6ml Collection Plate | 1 | 4 |
| 0.5ml Collection Plate | 1 | 4 |
| Silicon Seal Tape | 1 | 4 |
| Seal Film | 5 | 25 |
| Buffer ATL | 30 ml | 100 ml |
| Buffer AL | 30 ml | 100 ml |
| Buffer DW1 | 60 ml | 250 ml |
| Buffer GW2 | 50 ml | 2 x 100 ml |
| Proteinase K | 50 ml | 200 ml |
| Protease Dissolve Buffer | 5 ml | 15 ml |
| Buffer AE | 30 ml | 120 ml |
Storage and Stability
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 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 |
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:
TruScript Reverse Transcriptase is Available in Three Convenient Formats:
1. TruScript Reverse Transcriptase Kit (Cat.#54440)
This kit contains 5X RT Buffer and a vial of TruScript Enzyme Mix (200 units/µL). This enzyme can be used for reverse-transcription reactions with any user-supplied primers.
2. TruScript First Strand cDNA Synthesis Kit (Cat.#54420)
This is an all-in-one, ready-to-use product for simple set-up of reverse transcription of total RNA (both poly A- or non-poly A-containing transcripts).
The kit contains the 2x Reaction Mix and the TruScript Enzyme Mix. The 2x Reaction Mix contains a blend of buffer, nucleotides and primers (oligo dT and random hexamers) for effective cDNA synthesis from total RNA transcripts.
3. TruScript First Strand cDNA Synthesis Kit for mRNA (Cat.#54400)
This is an all-in-one, ready-to-use product for simple set-up of reverse transcription of messenger RNA (poly A-containing transcripts).
The kit contains the 2x Reaction Mix and the TruScript Enzyme Mix. The 2x Reaction Mix contains a blend of buffer, nucleotides and oligo dT primer for the effective cDNA synthesis from total RNA transcripts or enriched mRNA sample.
| TruScript Reverse Transcriptase | TruScript First Strand cDNA Synthesis Kit for mRNA | TruScript First Strand cDNA Synthesis Kit | |
| Kit contains only reverse transcriptase enzyme and buffer (no primers or other buffer components) | |||
| Your template is enriched mRNA | |||
| Your template is total RNA (poly A OR non-poly A-containing transcripts) | |||
| Kit contains oligo dT | |||
| Kit contains both oligo-dT primer and Random Hexamers | |||
| Kit contains reaction buffer with nucleotides and primer | with oligo-dT | with oligo-dT and random hexamers | |
| You have your own first strand synthesis primer | |||
| Your template is microRNA (using your own primers) | |||
| Cat.#54440 | Cat.#54400 | Cat.#54420 |
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TruScript First Strand cDNA Synthesis Kit (Cat.# 54420) – Kit Components
1. TruScript Enzyme Mix
2. 2x Reaction Mix
3. Nuclease-Free Water
4. Product Insert
Storage Conditions and Product Stability
Norgen’s TruScript Reverse Transcriptase and the 5x RT Buffer should be stored at -20°C. These reagents should remain stable for at least 1 year in their unopened containers.
| Component | Cat. 54440 (10,000 units) | Cat. 54420 (50 rxns) | Cat. 54410 (50 rxns) |
|---|---|---|---|
| TruScript™ Reverse Transcriptase (200 units/ μL) | 50 μL | – | – |
| 5x RT Buffer | 1 mL | – | – |
| TruScript™ Enzyme Mix | – | 100 μL | 100 μL |
| 2x Reaction Mix | – | 500 μL | – |
| 2x Reaction Mix for mRNA | – | – | 500 μL |
| Nuclease-Free Water | 1.25 mL | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 | 1 |
PEG3-(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane)-(Amino-Tri-(endo-BCN-PEG2-ethoxymethyl)-methane) is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The DBCO groups are commonly used for copper-free Click Chemistry reactions due to their strain promoted high energy. The hydrophilic PEG chain allows for increased water solubility.
PEG3-(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane)-(Amino-Tri-(endo-BCN-PEG2-ethoxymethyl)-methane) is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The DBCO groups are commonly used for copper-free Click Chemistry reactions due to their strain promoted high energy. The hydrophilic PEG chain allows for increased water solubility.