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 whole blood, tissue and culture cells.

Details

Specifications

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
• Pertinence – specially designed for isolating DNA from 3-10ml blood and related body fluids
• Wide applicability – handle a variety of liquid samples
• Economy – excellent cost effectiveness performance

Kit Contents

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

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:

Overview

• Purification and enrichment of intact exosomes from plasma, serum, urine, cell culture media and saliva in less than 30 minutes.
• Versatile sample input ranging from 250 µL to 30 mL
  • Urine Exosome Purification Mini Kit (250 µL – 1 mL urine)
  • Urine Exosome Purification Midi Kit (2 mL – 10 mL urine)
  • Urine Exosome Purification Maxi Kit (11 mL – 30 mL urine)
• Exosome purification is based on Norgen’s proprietary resin separating matrix through exosomes’ surface proteins.
• No precipitation reagents, overnight incubation, protease or coagulant treatments required
• No time-consuming ultracentrifugation, filtration or special syringes required
• Purify intact exosomes with a size ranging from 40-200 nm depending on sample input type
• Purified exosomes are compatible with functional studies.
• Purified exosomes are free from any protein-bound cell-free circulating RNA
• Purified exosomes are compatible with NanoSight® or Electron Microscopy for assessing the approximate exosome size range and concentration.
• Exosomal RNA can be extracted from the purified exosomes using Norgen’s Exosomal RNA Purification technology or any other RNA extraction method.

The Urine Exosome Purification Kits provide a fast, reliable and convenient method to purify and enrich for pure intact exosomes from different urine sample volumes ranging from 250 µL to 30 mL. These kits also allow for the purification of intact extracellular vesicles (EVs) from different urine sample volumes, and these EVs are ready for any downstream application. The purification is based on Norgen’s proprietary resin.

These kits provide a clear advantage over other available methods since they do not require any special instrumentation, ultracentrifugation, precipitation reagents or any protease treatments. More importantly, the purified exosomes will not be contaminated with any other RNA-binding proteins that may contaminate your exosomal RNA, which is essential if studying exosomal RNA gene expression.

NanoSight® Analysis

Exosomes enriched with Norgen’s Urine Exosome Purification Kits can be analyzed using NanoSight® for assessing the approximate exosome size range and concentration.

Exosomal RNA Analysis

Exosomal RNA can be isolated using Norgen’s Exosomal RNA Isolation Kit from exosomes enriched using Norgen’s Urine Exosome Purification Kits for gene expression analysis using RT-qPCR, microarray or NGS and for Biomarker discovery.

Details

Supporting Data

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Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. These kits are stable for 2 years after the date of shipment.

Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.

Description

Specifications

Cluster of differentiation 44 (CD44) is a glycoprotein receptor for hyaluronic acid, which plays a fundamental role in cellular adhesion, stromal binding, migration, and cell-cell interactions. Studies have suggested that the CD44-hyaluronate interaction is central to tumor invasiveness. Positive staining with Anti-CD44 is implicated in a multitude of different cancer types, including breast, prostatic, renal cell, colonic, hepatocellular, and genitourinary carcinomas, as well as Non-Hodgkin’s Lymphoma, metastatic melanoma, gastric cancer, and some soft tissue tumors. It has also been demonstrated that there is a positive correlation between tumor progression and increased expression of CD44v, a high molecular weight CD44 isoform that has been described in epithelial cells. Given the expression of CD44 in a wide range of cancers, the most practical application of CD44 immunostaining is its use in discriminating between urothelial transitional cell carcinoma in situ from non-neoplastic changes in the urothelium.