Overview

• Detection kits for WNV
• CE-IVD marked version available for in vitro diagnostic use
• Available in TaqMan format for analysis

West Nile Virus (WNV) belongs to the RNA virus family of Flaviviridae. It is known to spread primarily through arthropod vectors such as mosquitoes. The virus infects mainly birds but is also reported to infect humans as well as pets such as cats and dogs. In humans, the majority of the infections caused by WNV are asymptomatic. However, some individuals (the majority of the confirmed cases) could enter a second, febrile stage with flu-like symptoms – commonly known as West Nile Fever. In a more serious stage, the disease becomes neuroinvasive, causing meningitis or encephalitis. Such severe conditions could lead to mortality. As the symptoms of WNV are very similar to other common diseases but can be fatal at a severe stage, it is important to distinguish the virus early in the diagnosis, particularly at the molecular level.

WNV TaqMan RT-PCR Kit, 100 reactions

• Ready to use format, including Master Mix for the target and PCR control to monitor for PCR inhibition and validate the quality
• Specific Primer and Probe mix for the pathogen/virus/viroid of interest
• Primer and Probe mix
• Positive and negative control to confirm the integrity of the kit reagents

WNV TaqMan RT-PCR Probe/Primer Set and Controls, 100 reactions

• Specific Primer/Probe mix and Positive Control for the pathogen/virus/viroid of interest
• Nuclease-free water
• Can be used together with Norgen’s RT-PCR Master Mix (#28113) or customer supplied master mix

Details

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. TM44250 (100 preps)	Cat. TM44210 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
WNV Primer & Probe Mix	280 μL	280 μL
WNV Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Introduction

Hipure Stool RNA Kit is specially designed for stool RNA extraction. This kit is suitable for extracting high-purity microbial or host cell RNA from ≤0.1g stool samples. The kit adopts silica gel column purification technology and original solution system, which can effectively remove humic acid and other inhibitory factors in stool samples. The purified RNA can be directly used in RT-PCR, Northern hybridization and other experiments.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from 100-150mg stool sample
Applications	RT-PCR, Northern hybridization and other experiments
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Stool
Sample amount	100-150 mg
Elution volume	≥30μl
Time per run	≤50 minutes
Liquid carrying volume per column	100µg
Binding yield of column	800µl

Principle

The HiPure silica gel column uses a high binding ability glass fiber filter membrane as the substrate. Under the condition of high concentration of ionizing agent (such as Guanidinium chloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bonding and electrostatic and other physical factors, while protein or other impurities are not adsorbed and removed. The filter membrane that has adsorbed nucleic acids is washed to remove proteins and salts. Finally, low salt buffer solution (such as Buffer TE) or water can be used to wash out the nucleic acids adsorbed on the filter membrane. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.

The stool samples are homogenized in the lysis solution, further lysed in a high-temperature water bath, and RNA is released into the lysis solution. Chloroform extraction removes genomic DNA and impurities, transfer the supernatant to an alcohol free binding solution, purify RNA through a column, and finally elute RNA with RNase Free Water. The purified RNA can be directly used for experiments such as PCR, Southern hybridization, and enzyme digestion.

Advantages

• High purity – unique adsorbent for more efficient removal of inhibitors
• High concentration – maximum extraction of total RNA from stool samples
• Sensitive – RNA can be purified at the level of PG

Kit Contents

Contents	R418502	R418503
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	250
Glass Beads (0.1~0.6mm)	30 g	150 g
Buffer SPL	30 ml	140 ml
Buffer PHC	30 ml	140 ml
Buffer GRP	60 ml	250 ml
Buffer RW1	50 ml	250 ml
Buffer RW2 *	20 ml	2 x 50 ml
RNase Free Water	15 ml	30 ml

Storage and Stability

The kit components can be stored at room temperature (15–25°C) and are stable for 18 months under these conditions. At low temperatures, Buffer SPL may form precipitates, dissolve it by 55°C water bath. After receiving the product, Buffer PHC should be stored at 2-8°C.

Hipure Stool RNA Kit is specially designed for stool RNA extraction. This kit is suitable for extracting high-purity microbial or host cell RNA from ≤0.1g stool samples. The kit adopts silica gel column purification technology and original solution system, which can effectively remove humic acid and other inhibitory factors in stool samples. The purified RNA can be directly used in RT-PCR, Northern hybridization and other experiments.

DBCO-C5-NHS ester is an amine-reactive compound, which can be used to modify an amine-containing molecule in organic media. This reagent isn’t soluble in aqueous media. The extended 5-carbon atom spacer arm improves solubility in commonly used organic solvents including dichloromethane, chloroform, THF, and ethyl acetate, and it also improves derivatization efficiency and stability of conjugates. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research use only.

DBCO-C5-NHS ester is an amine-reactive compound, which can be used to modify an amine-containing molecule in organic media. This reagent isn’t soluble in aqueous media. The extended 5-carbon atom spacer arm improves solubility in commonly used organic solvents including dichloromethane, chloroform, THF, and ethyl acetate, and it also improves derivatization efficiency and stability of conjugates. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research use only.