Overview

• Detection kits for XMRV
• Available in TaqMan format for analysis

Xenotropic murine leukemia virus-related virus (XMRV) is a gammaretrovirus. The virus was first described in 2006 and has since been isolated from human biological samples. XMRV belongs to the family Retroviridae and the genus gammaretrovirus. It has a single-stranded RNA genome that replicates through a DNA intermediate. The virus gets its name due to its close relationship with the murine leukemia viruses (MuLVs). The viral genome is approximately 8100 nucleotides in length and is 95% identical with several endogenous retroviruses of mice. While gammaretroviruses have well-characterized oncogenic effects in animals, they have not been shown to cause human cancers. However, XMRV was recently discovered in human prostate cancers and is the first gammaretrovirus known to infect humans. In addition to prostate cancer, a possible association with chronic fatigue syndrome has been reported, however it has yet to be established whether XMRV is a cause of this disease.

The causal role of XMRV in cancer has yet to be established and the virus does not appear to be capable of transforming cells directly. In prostate cancer, XMRV protein has been found in tumour-associated but nonmalignant stromal cells, but not in the actual prostate cancer cells. This raises the possibility that the virus may support tumorigenesis. In other studies, XMRV proteins and nucleic acids were found in malignant cells.

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Supporting Data

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Storage Conditions and Product Stability
All kit components can be stored for 1 year after the date of production without showing any reduction in performance.

All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) 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.

Component	Cat. TM34750 (100 preps)	Cat. TM34710 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
XMRV Primer & Probe Mix	280 μL	280 μL
XMRV Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Overview

• Purification and enrichment of intact cell culture media exosomes for functional studies
• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias.
• Versatile cell culture media input volumes
• 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
• 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 Cell Culture Media Exosome Purification and RNA Isolation Kits constitute an all-in-one system for the purification of exosomes and the subsequent isolation of exosomal RNA from different cell culture media sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. The kit is 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. Moreover, the kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. 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.

Cell Culture Media Exosome Purification and RNA Isolation Mini Kit

For sample input volumes ranging from 5 mL to 10 mL.

Cell Culture Media Exosome Purification and RNA Isolation Midi Kit

For sample input volumes ranging from 10 mL to 20 mL.

Cell Culture Media Exosome Purification and RNA Isolation Maxi Kit

For sample input volumes ranging from 20 mL to 35 mL.

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Supporting Data

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Kit Specifications
Sample Type	Cell-Free Cell Culture Media
Sample Volume Range	20 mL to 35 mL
Size of RNA Purified	All sizes, including small RNA (<200 nt)
Elution Volume	50-100 μL
Time to Complete 10 Purifications	40 to 45 minutes
Average Yields*	Variable depending on specimen

* Please check page 4 of the product insert for Average Yields and Common RNA Quantification Methods

Storage Conditions and Product Stability
All buffers 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. 60700 (50 preps)	Cat. 60800 (25 preps)	Cat. 60900 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	1.5 mL	1.5 mL	1.5 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	18 mL	18 mL	18 mL
Elution Solution A	6 mL	6 mL	6 mL
Mini Filter Spin Column	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

Introduction

This product is suitable for extracting total viral nucleic acid from cell-free/low-content cell biological samples such as body fluids, serums, plasma, soaking solutions, tissue homogenate supernatant, and culture supernatant. The Purified DNA/RNA is used for RT-PCR and PCR detection.

Details

Specifications

Features	Specifications
Main Functions	IVD5412 precast kit for kingfisher Flex
Applications	RT-PCR，PCR，NGS
Products	Viral DNA / RNA, body cell DNA / RNA
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technolog
Process method	Manual or automatic
Sample type
Sample amount	200μl
Adaptive instrument	Nucleic acid extractor, pipetting workstation

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. DNA/RNA is released into the lysate. After adding magnetic particles and binding solution, DNA/RNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA/RNA was eluted by Nuclease Free Water.

Advantages

Kit Contents

Cat.No	Reagent	IVD5412-F-96
PK/Carrier RNA		1x 24 mg/Bottle
Protease Dissolve Buffer Blue		1.8 ml/Bottle
Tip		1
Sample Plate (DW Plate)	500µl Buffer MLB/Well	1
Wash 1 Plate (DW Plate)	500µl Buffer MW1/Well	1
Wash 2 Plate (DW Plate)	500µl Buffer CW/Well	1
Beads Plate (DW Plate)	500µl CW & 20µl MPN/Well	1
Elute Plate (KF Plate)	90µl NFW/Well	1

Storage and Stability

This kit is shipped and stored at room temperature and is valid for 12 months.

This product is suitable for extracting total viral nucleic acid from cell-free/low-content cell biological samples such as body fluids, serums, plasma, soaking solutions, tissue homogenate supernatant, and culture supernatant. The Purified DNA/RNA is used for RT-PCR and PCR detection.