Overview

• Detection kit for Listeria monocytogenes
• CE-IVDD marked in accordance with the European Commission Directive 98/79/EC.
• Research version available
• Available in TaqMan Probe format for analysis.

Listeria monocytogenes have emerged as significant foodborne pathogens that pose a serious public health problem. As the causative agent of Listeriosis, L. monocytogenes has the highest rate of fatality rate among all foodborne pathogens. L. monocytogenes is a facultatively intracellular, Gram-positive bacterium that could survive high and low temperatures, low pH. It is a rare causative agent of mastitis in cow. However, due to its ability to resist various food processing technologies as well as to grow at low temperature, L. monocytogenes is know to be associated with both raw and pasteurized milk, as well as dairy products such as cheese. As little as 1000 organisms may cause the disease with pregnant, new-born, and immunocompromised individuals the most susceptible.

Listeria monocytogenes TaqMan PCR Kit, 24 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

Details

Supporting Data

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Storage Conditions

The Listeria monocytogenes TaqMan PCR Kit Dx is shipped on dry ice. The components of the kit should be frozen upon arrival. If one or more of the components is not frozen when the kit is received, or if any of the components have been compromised during shipment, please contact Norgen Biotek for assistance. All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 3 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. DxTM30400 (24 rxns)
MDx TaqMan 2X PCR Master Mix Dx	550 µL
L. monocytogenes Primer Mix Dx	2 x 70 µL
L. monocytogenes Positive Control Dx – 200,000 copies/μL	50 µL
Nuclease-Free Water	2 x 1.25 mL
Product Insert	1

Introduction

This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from tissue / blood / body fluid / swab /dry blood spots
Applications	PCR, qPCR, southern bolt and virus detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Tissue, cell, blood, saliva, swab, blood spot, semen and other clinical samples
Sample amount	Solid tissue : 1-10mg, Anticoagulant blood : 200µl
Yield	1 – 15µg
Elution volume	≥20μl
Time per run	30 – 60 minutes
Liquid carrying volume per column	750µl
Binding yield of column	100µg

Principle

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 protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High quality DNA – meet a variety of downstream applications, including PCR, qPCR, enzyme digestion, hybridization, etc.
• Fast – without separation of leukocytes, organic extraction or ethanol precipitation
• Simple – all nucleic acids can be obtained by direct digestion
• Wide applicability – It can handle various liquid samples, animal tissues and cultured cells

Kit Contents

Contents	IVD3018
Purification Times	100
HiPure DNA Mini Columns I	100
2ml Collection Tubes	2 x 100
Buffer ATL	60 ml
Buffer AL	60 ml
Buffer GW1	44 ml
Buffer GW2	50 ml
Proteinase K	60 mg
Protease Dissolve Buffer	5 ml
Buffer AE	15 ml

Storage and Stability

Proteinase K 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 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.

Experiment Data

This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.

Introduction

The Kit is specially designed for simultaneous purification of genomic DNA and total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. Purified DNA/RNA is suitable for use in applications such as real-time PCR and Pyrosequencing.

Details

Specifications

Features	Specifications
Main FunctionsC	Co-isolation total RNA and DNA from FFPE tissue
Applications	RT-PCR, cDNA synthesis, PCR and second-generation sequencing, etc.
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Adaptive instrument	Nucleic acid extractor and pipetting workstation
Sample type	FFPE slice, FFPE puncture sample, embedded tissue
Sample amount	No more than six 10µm sections of 150 mm2 surface area or three 20µm sections of 150 mm2 surface area.

Principle

FFPE samples are incubated in an optimized lysis buffer, which results in the release of RNA and precipitation of DNA. After centrifugation, the RNA-containing supernatant and DNA-containing pellet are then processed separately to purify RNA and DNA. After adding magnetic particles and binding solution, DNA/RNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA/RNA was eluted by RNase Free Water.

Advantages

• Post digestion sorting – higher DNA and RNA yields
• Economy – the price is much lower than imported reagents

Kit Contents

Contents	IVD3026
Purification Times	200 Preps
MagBind Particles	9.0 ml
Proteinase K	180 mg
Protease Dissolve Buffer	10 ml
Buffer DPS	150 ml
Buffer FRL	40 ml
Buffer ATL	40 ml
Buffer AL	80 ml
Buffer BXW1*	110 ml
RNase Free Water	30 ml

Storage and Stability

Proteinase K and MagBind Particles 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 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.

The Kit is specially designed for simultaneous purification of genomic DNA and total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. Purified DNA/RNA is suitable for use in applications such as real-time PCR and Pyrosequencing.