Overview

• Detection kits for Legionella sp.
• CE-IVDD marked in accordance with the European Commission Directive 98/79/EC.
• Research version available

Legionella sp. TaqMan PCR Kit Dx, 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 Legionella sp. 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. DxTM64400 (24 rxns)
MDx TaqMan 2X PCR Master Mix	550 μL
Legionella sp. Primer & Probe Mix Dx	2 x 70 μL
Legionella sp. Positive Control Dx – 200,000 copies/μL	50 μL
Nuclease-Free Water	2 x 1.25 mL
Product Insert	1

Introduction

This product supplies a simple and rapid extraction of total RNA from tissue and culture cells samples. The kit is based on superparamagnetic particles purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction processis only 60 minutes. Purified RNA is ready for downstream applications such as RT-PCR, virus RNA testing and so on. MagPure LQ Kit buffers can be used for both manual extraction process and automatic nucleic acid extraction machines.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from tissue and cells and plant
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Adaptive instrument	Nucleic acid extractor, pipetting workstation
Sample type	Animal tissue, human tissue, cultured cells, lymphocytes, ordinary plant tissue
Sample amount	Cells: 1 x 107Animal tissue: ≤20mgPlant tissue: ≤100mgYeast cell: ≤5 x 106
Yield	5-100μg

Principle

The Kit combines the speed and efficiency of silica-based technology with the convenient handling of magnetic particles for purification of total RNA. Samples are lysed and RNA is purified from lysates in one step through its binding to the silica surface of the particles in the presence of a chaotropic salt. The particles are separated from the lysates using a magnet and DNA is removed by treatment with RNase-free DNase. The magnetic particles are efficiently washed, and RNA is eluted in RNase-free water.

Advantages

• High purity – OD 260 / 280 :1.9-2.0, OD 260 / 230 :1.5-2.0
• Economy – less than 50% of the price of Qiagen and other imported products
• Automatic – extraction without manual participation, saving time and effort
• Universal – suitable for various animal tissues, cultured cells and conventional plant fungal tissues

Kit Contents

Contents	R662201	R662202	R662203
Purification Times	48 Preps	96 Preps	5 x 96 Preps
MagPure RNA Particles	1.7 ml	4.0 ml	18 ml
Proteinase K	24 mg	50 mg	240 mg
Protease Dissolve Buffer	1.8 ml	5 ml	15 ml
DNase I	600 μl	2 x 600 μl	10 x 600 μl
DNase Buffer	30 ml	40 ml	200 ml
RTL Lysis Buffer	40 ml	80 ml	400 ml
Buffer MCB*	18 ml	30 ml	150 ml
Buffer MW1*	44 ml	66 ml	2 x 220 ml
Buffer RW2*	20 ml	50 ml	2 x 100 ml
RNase Free Water	10 ml	30 ml	120 ml

Storage and Stability

MagPure RNA Particles and Proteinase K should be stored at 2–8°C upon arrival. DNase I shouldbe stored at -20°C. However, short-term storage (DNase I up to 1 weeks, MagPure RNA Particles and Proteinase K up to 8 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 atleast 18 months under these conditions.

This product supplies a simple and rapid extraction of total RNA from tissue and culture cells samples. The kit is based on superparamagnetic particles purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction processis only 60 minutes. Purified RNA is ready for downstream applications such as RT-PCR, virus RNA testing and so on. MagPure LQ Kit buffers can be used for both manual extraction process and automatic nucleic acid extraction machines.

The Bisulfite Sequencing Library Prep Kit (illumina platform) was developed for construction of NGS libraries using bisulfite treated DNA (50 ng – 500 ng) as input. DNA methylation is an epigenetic mechanism known to play a critical role in gene regulation and genomic imprinting by blocking transcription factor access to promoters and enhancers. Bisulfite sequencing is a popular technique in biomedical research based on C to T conversion under the treatment of sodium bisulfite.

Recently, NGS became a powerful tool to identify the DNA methylation status at the whole genome level with single-base resolution. However, it is well known that bisulfite treatment of the NGS libraries causes tremendous damage to the libraries.

Bisulfite-Seq kit comparison

In the case of the regular bisulfite seq library preparation (library prep before bisulfite conversion), the DNA shearing equipment and the expensive methylated adaptors are required. In addition, the subsequent bisulfite conversion causes tremendous DNA damage to the constructed libraries.

BioDynami has developed a unique library prep technology to solve the problems. The technology uses bisulfite treated DNA as input to avoid the significant library loss caused by bisulfite conversion. Furthermore, DNA shearing step and expensive methylated adaptors are not required with our kit. The DNA polymerase in the kit has high-fidelity amplification ability and uracil tolerance which is ideal for amplification of bisulfite sequencing libraries. The final library is strand specific.

Bisulfite Sequencing Library Prep Kit Workflow

Three index types are available for the kit:

Non-index (Cat.# 30091): Libraries do not have index.

Index (Cat.# 30092): Each primer contains a unique barcode sequence of 6 bases to identify the individual library. Library multiplexing capacity is up to 48 samples. Index information can be downloaded here.

Unique dual index (Cat.# 30093): The multiplexing of bisulfite sequencing library is up to 96 samples with unique dual indexes. We used a Four-Base Difference Index System to generate indexes that have at least 4 bases different from each other in the 8-base index. The index primers remove NGS errors including index cross-contamination, index hopping, reads mis-assignment etc. Index information can be downloaded here.

Kit advantages

• • • Easy and Quick protocol
      • Easy: Hands-on time only 10 minutes
      • Quick: Total protocol time around 1.5 hours
    • Simple workflow: Less steps
    • Directional library
    • Save magnetic beads more than 50%
    • Guaranteed high Bisulfite sequencing library conversion efficiency
    • Bisulfite treated DNA as input: From 50 ng to 500 ng

The Bisulfite Sequencing Library Prep Kit (illumina platform) was developed for construction of NGS libraries using bisulfite treated DNA (50 ng – 500 ng) as input. DNA methylation is an epigenetic mechanism known to play a critical role in gene regulation and genomic imprinting by blocking transcription factor access to promoters and enhancers. Bisulfite sequencing is a popular technique in biomedical research based on C to T conversion under the treatment of sodium bisulfite.