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 is suitable for extracting RNA  from 1ml anticoagulant blood (fresh or frozen blood/cryopreservation blood), lymphocytes, buffy coat, bone marrow, cultured cells, etc using Magzol reagents. It can specially extract high quality RNA from frozen blood samples.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from 1ml anticoagulant blood (fresh or frozen blood/cryopreservation blood), lymphocytes, buffy coat, bone marrow, cultured cells, etc using Magzol reagents.
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	anticoagulant blood, lymphocytes, buffy coat, bone marrow, cultured cells
Sample amount	1ml whole blood (fresh or frozen blood)

Principles

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested by lysis buffer and protease, and RNA/DNA is released into the lysis buffer. Add binding solution and magnetic particles to adsorb RNA/DNA, while proteins are not adsorbed and removed. The particles adsorbed with DNA/RNA are washed with washing buffer to remove proteins and other impurities, then washed with ethanol to remove salt, and finally digested with DNase to remove DNA. RNA is recovered by adding binding solution, and finally the RNA is eluted with low salt buffer. The eluted RNA can be directly used for experiments such as RT-PCR, NGS and virus detection.

Kit Contents: Bottle

Contents	R661301	R661302	R661303
Purification Times	48 Preps	96 Preps	480 preps
DNase I	600 μl	2 x 600 μl	10 x 600 µl
DNase Buffer	20 ml	30 ml	150 ml
MagPure Particles N	2.5 ml	5.0 ml	28 ml
MagZol 3BD	65 ml	140 ml	3 x 200 ml
Buffer ALB2	40 ml	60 ml	350 ml
Buffer BCP2	10 ml	15 ml	80 ml
Buffer MW2*	20 ml	50 ml	2 x 100 ml
RNase Free Water	10 ml	20 ml	60 ml

Storage and Stability

DNase I should be shipped with ice pack or dry ice and stored at -20°C upon arrival. MagPure Particles N, MagZol 3BD and Buffer BCP2 should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage 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 conditions.

This product is suitable for extracting RNA from 1ml anticoagulant blood (fresh or frozen blood/cryopreservation blood), lymphocytes, buffy coat, bone marrow, cultured cells, etc using Magzol reagents. It can specially extract high quality RNA from frozen blood samples.

Introduction

The HiPure FastFilter Plasmid DNA Maxi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. This system also includes a special filter cartridge which replaces the centrifugation step following alkaline lysis. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 1000 µg high copy number plasmid DNA or 50-500 µg low copy number plasmid DNA can be purified from 200mL overnight culture.

Details

Specifications

Features	Specifications
Main Functions	Isolation up to 1mg plasmid DNA from 200ml bacterial culture
Applications	Enzyme digestion, sequencing, PCR and labeling,  etc.
Purification method	Maxi spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Conventional plasmid, plasmid less than 30KB
Sample amount	100-200ml
Yield	0.4-1mg
Elution volume	≥500μl
Time per run	≤60 minutes
Liquid carrying volume per column	20ml
Binding yield of column	1mg

Principle

The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
• Fast – it takes only 15-60 minutes to complete the isolation
• Economy – high cost performance

Kit Contents

Contents	P101402	P101403
Purification Times	10 Preps	50 Preps
RNase A	20 mg	40 mg
Buffer P1	140 ml	2 x 350 ml
Buffer P2	140 ml	2 x 350 ml
Buffer LEN3	70 ml	350 ml
Buffer GBT	120 ml	550 ml
Buffer PW1	60 ml	300 ml
Buffer PW2*	50 ml	4 x 100 ml
Elution Buffer	20 ml	120 ml
HiPure DNA Maxi Columns III	10	50
Lysate Clear Maxi Syringe	10	50
50 ml Collection Tubes	20	100

Storage and Stability

The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. After addition of RNase A，Buffer P1 is stable for 6 months when stored at 2-8°C.

Purchase Guide

Name	CAT NO	Features	Sampleamount	Elutionvolume	Yield
HiPure Plasmid Mini Kit	P1001	Classic, Regular application	1-5ml LB	30-50μl	5–35µg/5ml
HiPure Plasmid Mini Kit II	P1002	Classic, Regular application	5-15ml LB	75-100μl	20-80µg/15ml
HiPure Plasmid Plus 96 Kit	P1006	Classic, Regular application	1-1.5ml LB	70-150μl	1-15μg/1ml
HiPure Fastfilter Plasmid Midi Kit	P1013	Fast	30-50ml LB	0.3-0.8ml	50–250µg/50ml
HiPure Fastfilter Plasmid Maxi Kit	P1014	Fast	100-200ml LB	0.5-1.5ml	0.4–1mg/200ml
Low endotoxin/Endotoxin-free Plasmid
HiPure Plasmid EF Maxi Kit	P1156	Classic method, General for all kinds of vectors	100-200ml LB	≥0.7ml	200–1500µg
HiPure Plasmid EF Mini Kit	P1154	Recommend for low copy vector, Thoroughly remove RNA	5-15ml LB	≥60μl	10–80µg
HiPure Plasmid EF 96 Kit	P1157	Recommend for low copy vector, Thoroughly remove RNA	1-5ml LBx96	≥75μl	30µg
HiPure Plasmid EF Midi Kit	P1231	Recommend for High copy vector, Low elution volume, High concentration	25-50ml LB	≥100μl	10-500μg
HiPure Plasmid EF Mega Kit	P1116	Recommend for High copy vector(>3μg/ml)	500ml LB	≥2ml	1-10mg

For any technical problems or customized products, please contact us.

F&Q about Endotoxin-free Plasmid Extraction Kit — P1156 ←click here

The HiPure FastFilter Plasmid DNA Maxi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. This system also includes a special filter cartridge which replaces the centrifugation step following alkaline lysis. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 1000 µg high copy number plasmid DNA or 50-500 µg low copy number plasmid DNA can be purified from 200mL overnight culture.