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Under the tab \"Downloads\" you will find an Application Note showing data using the LightCycler® Multiplex DNA Master and the LightCycler® Multiplex RNA Virus Master on the following instruments:
Applied Biosystems QuantStudio 7 Flex RealTime PCR System
Applied Biosystems 7500 RealTime PCR System
Applied Biosystems 7900HT Fast RealTime PCR System
Applied Biosystems StepOnePlusRealTime PCR System
and BioRad CFX96 RealTime PCR Detection System.", "Country": "XG", "Code": "Protocols - Help Corner", "Name": "Protocols - Help Corner" }, { "Language": "en", "Value": "
How this Product Works
The LightCycler® Multiplex RNA Virus Master consists of 3 different vials:
  • Vial 1: RT Enzyme Solution (Reverse Transcriptase)
  • Vial 2: RT-qPCR Reaction Mix
  • Vial 3: Water, PCR Grade
The separate vial of Reverse Transcriptase makes it possible to prepare a RT-qPCR reaction mix for running RT-minus controls (lacking reverse transcriptase). This is important for verifying whether the obtained results are derived from RNA transcripts or from residual (contaminating) genomic DNA. The kit also provides sufficient vials of Water, PCR Grade to ensure that fresh (unopened) vials can be used. This minimizes the risk of contamination of RT-qPCR reaction mixes by RNases and other substances.
The Reverse Transcriptase provided in this kit is a recombinant reverse transcriptase with higher stability than native reverse transcriptase. This feature allows for higher reverse transcription temperatures of up to +55°C. This Roche recombinant Reverse Transcriptase also has lower affinity for DNA than other commonly used reverse transcriptases.
The resulting RT-qPCR reaction mix also has AptaTaq DNA Polymerase, nucleotides, and additives, ensuring a hot start amplification system with high specificity. The mix contains an optimized concentration of MgCl2, eliminating the need for additional adjustments. For greater convenience, the RT-qPCR mixture, including Reverse Transcriptase, can be used for both RNA and/or DNA templates in parallel.", "Country": "XG", "Code": "Principle", "Name": "Principle" }, { "Language": "en", "Value": "
Standard Laboratory Equipment
  • Nuclease free pipette tips
  • 1.5 ml RNase-free microcentrifuge tubes to prepare master mixes and dilutions.
  • To minimize risk of RNase contamination, autoclave all vessels.
    Wear gloves at all times.
For RT-qPCR
  • Real-Time PCR systems such as the LightCycler® 480 Instrument*, LightCycler® 96 Instrument*, LightCycler® Nano Instrument, or the LightCycler® 2.0 Instrument*
  • LightCycler®480 Multiwell Plate 96* or 384*
  • LightCycler® 8-Tube Strips*
  • LightCycler® 8-Tube Strip Adapter Plate*
  • LightCycler® Capillaries (20 μl)*
  • Standard swinging-bucket centrifuge with rotor for multiwell plates
  • Real-Time PCR systems such as Applied Biosystems QuantStudio™ 7 Flex Real-Time PCR System,
    Applied Biosystems 7500 Real-Time PCR System,
    Applied Biosystems 7900HT Fast Real-Time PCR System,
    Applied Biosystems StepOnePlus™Real-Time PCR System,
    and Bio-Rad CFX96™ Real-Time PCR Detection System
For RT-qPCR Primer and Probe Design
  • Universal ProbeLibrary Assay Design Center at www.universalprobelibrary.com
  • To design and order RT-qPCR assays and panels: www.realtimeready.roche.com
Optional For Virus RNA Purification
  • MagNA Pure 96 Instrument* including disposables
  • MagNA Pure 96 Internal Control Tube*, optional
  • MagNA Pure 96 DNA and Viral NA Kit, Large Volume* or
  • MagNA Pure 96 DNA and Viral NA Kit, Small Volume*

Alternatively, use other nucleic acid purification instruments and manual sample preparation products such as:
  • MagNA Pure LC 2.0 Instrument with MagNA Pure LC Total Nucleic Acid Isolation Kit High - Performance*
  • MagNA Pure Compact Instrument with MagNA Pure Compact Nucleic Acid Isolation Kit I*
  • High Pure Viral Nucleic Acid Kit*
", "Country": "XG", "Code": "Additional Equipment and Reagent Required", "Name": "Additional Equipment and Reagent Required" }, { "Language": "en", "Value": "
Vial / BottleCapLabelFunction / DescriptionCatalog NumberContent
1blueLightCycler® Multiplex RNA Virus Master,
RT Enzyme Solution, 200x conc.
Contains Reverse Transcriptase.06 754 155 001 1 vial,
28 μl
07 083 173 001 5 vials,
28 μl each
2redLightCycler® Multiplex RNA Virus Master,
RT-qPCR Reaction Mix, 5x conc.
Contains RT-qPCR Reaction Buffer, AptaTaq Polymerase, dATP, dCTP, dGTP, dUTP, MgCl2, and proprietary additives.06 754 155 001 1 vial,
880 μl 
07 083 173 001 5 vials,
880 μl each
3colorlessLightCycler® Multiplex RNA Virus Master,
Water, PCR Grade
 To adjust the final reaction volume.06 754 155 001  3 vials,
1 ml each 
07 083 173 001 15 vials,
1 ml each
", "Country": "XG", "Code": "Content", "Name": "Content" }, { "Language": "en", "Value": "Optimized for multiplex PCR of 3 or more targets and an internal control RNA, this 5x master mix is especially suited for the fast and efficient detection of virus parameters in human sample materials. The robust formulation allows multiplex one-step RT-qPCR detection of a multitude of viral targets without the need for reaction condition optimization or adaptation. DNA and RNA targets can be run with the same PCR protocol on the LightCycler® 480, LightCycler® 96, and LightCycler® Carousel-based Instruments.", "Country": "XG", "Code": "Applications", "Name": "Applications" }, { "Language": "en", "Value": "A triplex protocol using 45 cycles including a 10-minute reverse transcription step requires 65 minutes using the LightCycler® 480 System or 67 minutes using the LightCycler® 96 System.
 
", "Country": "XG", "Code": "Assay Time", "Name": "Assay Time" } ] } } ] }

LightCycler® Multiplex RNA Virus Master

Easy-to-use reaction mix for one-step RT-qPCR using the LightCycler® 480, LightCycler® 96, LightCycler® Nano, or the LightCycler® 2.0 Real-Time PCR Systems

LightCycler<sup><sup>®</sup></sup> Multiplex RNA Virus Master

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