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 (fresh or frozen), plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from 200ul Whole Blood
Applications
PCR, southern bolt and virus detection, etc
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Whole Blood (fresh or frozen), serum, plasma, milk, saliva, and other liquid samples and cultured cells
Sample amount
<200μl whole blood or other liquid samples, <5*106 lymphocytes or Culture CellsNon mammalian animals that have nucleus in red blood cells (rich in DNA, such as birds and fish) : 5~20μl whole blood at a time
Elution volume
≥20μl
Time per run
≤30 minutes
Liquid carrying volume per column
800µl
Binding yield of column
100µg
Principle
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
D311102
D311103
Purification Times
50
250
HiPure DNA Mini Columns I
50
2 x 125
2ml Collection Tubes
100
5 x 100
Buffer AL
15 ml
60 ml
Buffer DW1
30 ml
150 ml
Buffer GW2*
12 ml
50 ml
Proteinase K
24 mg
120 mg
Protease Dissolve Buffer
1.8 ml
10 ml
Buffer AE
15 ml
60 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.
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:
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.
Background
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.
EXTRAClean Plasma/Serum RNA Purification Mini Kit
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.
EXTRAClean Plasma/Serum RNA Purification Midi Kit
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.
EXTRAClean Plasma/Serum RNA Purification Maxi Kit
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.
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.
Lyophilized format designed for room temperature shipping
Available in TaqMan format for analysis
Norgen’s EBV TaqMan PCR Lyophilized Kit is designed for the detection of EBV specific DNA in a real-time PCR based on the use of TaqMan® technology. The lyophilized format is designed to ship the kit at ambient temperature.
Norgen’s EBV TaqMan Lyophilized Probe/Primer and Control Set is designed for the detection of EBV specific DNA in a real-time PCR based on the use of TaqMan® technology. The lyophilized format is designed to ship the kit at ambient temperature.
All kit components should be stored at -20°C upon arrival.
Once reconstituted, repeated thawing and freezing (>2 times) 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.
Each kit is provided with 4 tubes of 2X PCR Master Mix and each tube is enough to run 25 reactions. It is not necessary to reconstitute all Master Mix tubes at once. The Master Mix tubes can be reconstituted as and when needed.
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
