Overview

• Detection kits for H5N1
• Available in TaqMan format for analysis

Influenza virus infection of birds, humans and other animals is a major public health problem worldwide. Influenza viruses are classified as either type A, B or C based on differences in their nucleoproteins and matrix proteins. The type A viruses are the most virulent human pathogens among the three influenza types and cause the most severe disease and epidemics. The different types can be further classified into subtypes based on antigenic differences in two surface glycoproteins; hemagglutinin and neuroamidase. All known subtypes of influenza A can be found in birds (H1-H16, N1-N9), while a limited number of the subtypes have been found in humans (H1-H3, N1 and N2). However, over the past few years, various subtypes of Influenza A viruses, including H5N1, have been reported to infect humans (WHO, 2006). In addition, the coexistence of human influenza viruses and avian influenza viruses may provide an opportunity for genetic material to be exchanged between these viruses. This could potentially create a new virulent influenza strain that is easily transmissible and lethal to humans (Food Safety Research Information Office, 2006). Thus, there is the need for sensitive diagnostic tests to allow for the rapid and early detection of these H5 influenza virus infections, to help reduce the risk of epidemics or pandemics in both animals and humans.

H5N1 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

H5N1 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

For research use only and NOT intended for in vitro diagnostics.

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. TM35450 (100 preps)	Cat. TM35410 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
H5N1 Primer & Probe Mix	280 μL	280 μL
H5N1 Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Introduction

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 tissue and culture cells.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from <25 mg tissue, culture cells, FTA card
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	Animal tissue or cultured cells
Sample amount	Animal tissue : <25mg, Cultured cells : <5 x 106
Elution volume	≥30μl
Time per run	30 – 60 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

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 proteinis not adsorbed and is removed with filtration. After washing proteins andother impurities, Nucleic acid was finally eluted with low-salt buffer (10mmTris, pH9.0, 0.5mm EDTA).

Advantages

• Good repeatability – suitable for extracting high-yield DNA from different types of tissue samples
• High purity – can be used for downstream applications such as multiplex and quantitative PCR
• Fast – simplified process, extracting several samples in 20 minutes (after digestion)
• Safety – no phenol orchloroform extraction, no alcohol precipitation

Kit Contents

Contents	D312102	D312103
Purification Times	50 Preps	250 Preps
Buffer ATL	15 ml	65 ml
Buffer DL	15 ml	65 ml
Buffer GW1	22 ml	110 ml
Buffer GW2	12 ml	50 ml
RNase A	10 mg	50 mg
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	5 ml	15 ml
Buffer AE	15 ml	60 ml
HiPure gDNA Mini Columns	50	2 x 125
2 ml Collection Tubes	100	5 x 100

Storage and Stability

RNase A and Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

Purchase Guide

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 tissue and culture cells.

Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Primer and probe mix (150 reactions)
Reverse Transcription, target specific primers (RNA genome viruses only)
Copy number standard curve (sufficient for multiple standard curves)
Internal extraction control – Read through VIC channel*
Endogenous control (150 tests)
RNAse/DNAse free water
*alternative fluorophores available on request