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.

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 kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. RNA purified using the HiPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA (not include miRNA) from 20mg tissue, 150mg plant, 5 x 106 cell using two columns (gDNA removed column)
Applications	RT-PCR, qRT-PCR, Northern hybridization, second generation sequencing
Purification method	Mini spin column
Purification technology	Silica technology, DNA filtration technology
Process method	Manual (centrifugation or vacuum)
Sample type	Animal soft tissue, cultured cells, lymphocytes, simple plant tissue
Sample amount	Cells:≤1 x 107 Animal tissue sample: 1-20 mgPlant tissue: 50-150 mg
Yield	2-100μg
Elution volume	≥50μl
Time per run	≤25 minutes（1-24 samples）
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

The Kit isolates total RNA from up to 10⁷ cells or 20 mg tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 25 mins. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column, ethanol is added to the flow-through, and the sample is applied to a RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit.

Advantages

• Efficient removal of DNA – unique genomic DNA removal column without DNase treatment
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – several samples can be extracted in 25 minutes by column method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG

Kit Contents

Contents	R411102	D411103
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns	50	250
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	100	2 x 250
Buffer RLC	50 ml	200 ml
Buffer RW1	50 ml	200 ml
Buffer RW2*	12 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

HiPureKit can be stored dry at room temperature (15-25°C) and are stable for at least18 months under these conditions. During shipment, crystals or precipitationmay form in the Buffer RLC. Dissolve by warming buffer to 37°C.

This kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. RNA purified using the HiPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Introduction

This kit is used for extracting total pathongen nucleic acid from low cell content biological samples such as body fluid, serum, plasma, immersion solution, tissue homogenate, culture, etc. The purified DNA/RNA is ready for downstream clinical in vitro detection such as Real Time PCR, biochip analysis, NGS and other related detection.

Details

Specifications

Features	Specifications
Main Functions	Extract pathogen DNA/RNA from whole blood, body fluid, serum, plasma, sputum, immersion solution, tissue homogenate supernatant, etc.
Applications	RT-PCR，PCR，NGS
Products	Pathogen DNA and RNA
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Whole blood, plasma, serum, immersion solution, tissue homogenate supernatant
Sample amount	200μl
Elution volume	≥30μl
Time per run	30 mins
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA/RNA is released into the lysate. Transfer to an adsorption plate and filter column. DNA/RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, DNA / RNA is finally eluted with low-salt buffer (10 MmTris, pH 8.0).

Advantages

• Fast – column purification, several samples can be finished in 30 mins
• High quality – high purity total RNA/DNA can be directly used in various sensitive downstream applications
• Safe – no phenol chloroform extraction required
• Sensitive – DNA/RNA can be recovered at the level of PG

Kit Contents

Contents	IVD4179
Purification Times	50 Preps
HiPure Viral Mini Column	50
2ml Collection Tubes	100
2ml Bead Tubes	50
Protease K	50 mg
Protease Dissolve Buffer	5 ml
DNase I (Powder)	10 mg
DNase Buffer	6 ml
Buffer CLB	100 ml
Buffer SDS	5 ml
Reagent DX	1.5 ml
Buffer ACL	30 ml
Buffer VHB*	22 ml
Buffer RW2*	20 ml
Buffer AVE	15 ml

Storage and Stability

Proteinase K and DNase I (Powder) should be stored at 2–8°C upon arrival. However, short-term storage (DNase I up to 1 week, Proteinase K up to 8 weeks) at room temperature (15–25°C) does not affecttheir performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for 18 months under these conditions.

This kit is used for extracting total pathongen nucleic acid from low cell content biological samples such as body fluid, serum, plasma, immersion solution, tissue homogenate, culture, etc. The purified DNA/RNA is ready for downstream clinical in vitro detection such as Real Time PCR, biochip analysis, NGS and other related detection.