Overview

• Ensure optimal results for sensitive applications like NGS
• Up to a tenfold increase in microRNA mapping during sequencing runs to reduce costs
• The purified exosomal RNA is free from any circulating RNA-binding proteins.
• No phenol extractions, Proteinase K treatment, nor carrier RNA.
• No time-consuming ultracentrifugation, filtration nor special syringes are required.
• Concentrate isolated RNA into a flexible elution volume ranging from 50 μL to 100 μL.
• Purification is based on EXTRAClean spin column chromatography that uses Norgen’s proprietary separation matrix.

Norgen’s EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Kit constitutes an all-in-one system for the purification of exosomes and the sequential isolation of RNA and free-circulating RNA from different plasma/serum sample volumes ranging from 50 μL and up to 10 mL. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS. The kit is designed to isolate all sizes of RNA, including microRNA. The kit provides a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or any protease treatments. Moreover, the kit allows 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. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA can be used in a number of downstream applications including real time PCR, NGS application, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Details

Supporting Data

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Kit Specifications (Spin Column)
Minimum Plasma Input	50 uL
Maximum Plasma Input	1 mL
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

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.

Description

As this is a 2 gene kit, we recommend purchase of 2 of the accompanying RT-qPCR master mix reagent: oasig Lyophilised OneStep RT-qPCR Master Mix 150 reactions.

Norovirus is known to cause acute gastroenteritis. It is a small (27-38 nm), round, nonenveloped RNA virus belonging to the Caliciviridae family and is responsible for over 80% of non-bacterial outbreaks of gastroenteritis in the world. It affects individuals of all ages, with a distinct seasonal link to winter. It has a genome of 7.6 kb that is positive sense and has a single stranded linear confirmation. It encodes a major structural protein (VP1) of about 58 to 60 kDa and a minor capsid protein (VP2). Transmission occurs predominantly through ingestion of contaminated water, food and airborne transmission, as well as contact with contaminated surfaces. The ease with which norovirus is transmitted and the low infectious dose required to establish an infection results in extensive outbreaks in numerous environments, such as hospitals, hotels and schools. There is no antiviral drug available to treat this infection and little is known about its pathogenicity. However, it has been observed that the virus can be taken up by enterocytes where translation of viral nonstructural proteins can occur; it damages and alters intestinal microvilli, leaving them blunt and broadened, thus inhibiting absorption; it causes crypt cell hyperplasia and also leads to apoptosis of enterocyctes. An incubation period of 24-48 hours is usual. Infection is characterized by the acute onset of nausea, vomiting, abdominal cramps, aching limbs, raised temperature and diarrhoea that generally last for about 48 hours. However, more severe and prolonged infection may be observed in children and the elderly. There are five recognized norovirus genogroups, of which three (GI, GII, and GIV) are known to affect humans and, since 2002, variants of the GII.4 genotype have been the most common cause of norovirus outbreaks. There have been 31 different genotypes identified within the genogroups, with a wide degree of genetic variability present even within each genotype.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings

Introduction

This product provide fast and easy methods for purification of total DNA from whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells. There is no need to use toxic phenol chloroform extraction or time-consuming alcohol precipitation. The extraction process finish in 40 minutes. Purified DNA includes genomic DNA, mitochondrial DNA, viral DNA (e.g. HBV), or DNA from other parasitic microorganisms. The obtained DNA can be directly used in PCR, viral DNA detection and other experiments.

This kit can use on manual protocol or 16/32/48 channel automated extraction system. No need to pause for the addition of binding solution (not applicable to 96/192 instruments)

Details

Kit Contents

Contents	D631001C	D631002C	D631003C
Purification Times	48	96	480
MagPure Particles	1.7 ml	3.5 ml	16 ml
Proteinase K	24 mg	50 mg	220 mg
Protease Dissolve Buffer	1.8 ml	3 ml	15 ml
Buffer MLA	30 ml	70 ml	300 ml
Buffer DW1	30 ml	70 ml	300 ml
Buffer EW	60 ml	120 ml	2 x 300 ml
Buffer MWX1	30 ml	70 ml	300 ml
Elution Buffer	10 ml	20 ml	60 ml

Prefilled Kit Contents

Product	Contents and volume	D6310-TL-06C	D6310-S-48
Proteinase K		50 mg	24 mg
Protease Dissolve Buffer		3 ml	1.8 ml
TL-Tip		12 pcs	24 pcs
Tip bottom plate/ Tip base reagent strip	Row 1/7：600μl Buffer MLA	6 plates	48 strips
	Row 2/8：600μl Buffer MWX1
	Row 3/9：600μl Buffer DW1
	Row 4/10：30μl MagPure Particles 600μl Buffer EW
	Row 5/11：600μl Buffer EW
	Row 6/12：100μl Elution Buffer

Storage and Stability

Proteinase K Powder 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 condition.

This product provide fast and easy methods for purification of total DNA from whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells. There is no need to use toxic phenol chloroform extraction or time-consuming alcohol precipitation. The extraction process finish in 40 minutes. Purified DNA includes genomic DNA, mitochondrial DNA, viral DNA (e.g. HBV), or DNA from other parasitic microorganisms. The obtained DNA can be directly used in PCR, viral DNA detection and other experiments.