Overview

• Purification and enrichment of intact plasma/serum exosomes for functional studies
• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias
• Versatile plasma/serum input volume
• No phenol extractions, Proteinase K treatment, nor carrier RNA required
• No time-consuming ultracentrifugation, filtration nor special syringes required
• No precipitation reagents, nor overnight incubation required
• Compatible with plasma/serum from most species
• Pure exosomes are purified and are free from any other RNA-binding proteins
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin
• The purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services

Norgen’s Plasma/Serum Exosome and RNA Isolation Kits constitute all-in-one systems for the purification of exosomes and the sequential isolation of exosomal RNA from different plasma/serum sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Norgen’s Plasma/Serum Exosome and RNA Isolation Mini Kit

For sample volumes ranging from 50 µL to 1 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.

Norgen’s Plasma/Serum Exosome and RNA Isolation Midi Kit

For sample volumes ranging from 1 mL to 4 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.

Norgen’s Plasma/Serum Exosome and RNA Isolation Maxi Kit

For sample volumes ranging from 4 mL to 10 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.

Details

Supporting Data

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Kit Specifications
Minimum Plasma/Serum Input	1 mL
Maximum Plasma/Serum Input	4 mL
Size of Exosomes Purified	40 nm – 150 nm
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume	50-100 μL
Time to Complete 10 Purifications	35 – 40 minutes
Average Yields	Variable depending on specimen

*Please check page 4 of the product insert for the average yields and the 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.

Important Note
This kit is suitable for the purification of exosomes 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.

Component	Cat. 58300 (50 preps)	Cat. 58500 (25 preps)	Cat. 58600 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	8 mL	2 x 8 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	20 mL	20 mL	20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	38 mL	18 mL	18 mL
Elution Solution A	6 mL	6 mL	6 mL
Mini Filter Spin Columns	50	25	15
Mini Spin Columns	50	25	15
Collection Tubes	50	25	15
Elution Tubes (1.7 mL)	100	50	30
Product Insert	1	1	1

DNA amplification in a flexible liquid format. Contains all the enzymes and reagents necessary for the amplification of DNA, the user need only supply primers, dNTPs and template. See manual for more information. Click to order oligonucleotides.

Perfect for:

• End-point gel electrophoresis DNA detection
• down-stream applications (e.g. sub-cloning)
• facilitating use of different RPA reaction volumes, or variation of component ratios.

Product code: TALQBAS01

DNA amplification in a flexible liquid format. Contains all the enzymes and reagents necessary for the amplification of DNA, the user need only supply primers, dNTPs and template. See manual for more information. Click to order oligonucleotides.

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 (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.

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: