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LightCycler® 480 Probes Master

Ready-to-use hot start reaction mix for probe-based real-time PCR using the LightCycler® 480 Instruments.

For life science research only. Not for use in diagnostic procedures.

Product No. Pack Size
04707494001 500 x 20 µL reactions
Product No. Pack Size
04887301001 5000 x 20 µL reactions (10 x 5 mL)
Product No. Pack Size
04902343001 5000 x 20 µL reactions (1 x 50 mL)

To order these products, please contact your Roche order management team.

LightCycler® 480 Probes Master is a ready-to-use reaction mix specifically developed for the hydrolysis probe detection format in multiwell plates on the LightCycler® 480 Instrument. It contains FastStart Taq DNA Polymerase for hot start PCR, which significantly improves the specificity and sensitivity of PCR by minimizing the formation of nonspecific amplification products.

 

  • Save time with a convenient, ready-to-use 2x-concentrated hot start master mix.
  • Minimize pipetting steps and contamination risk.
  • Perform sensitive, specific, and quantitative PCR or RT-PCR using the LightCycler® 480 Instruments.
  • Eliminate time-consuming MgCl2 titration.
  • Achieve consistent, high-quality performance with the LightCycler® 480 Instrument and Multiwell Plates.
  • Establish new qPCR assays and get results quickly by combining LightCycler® 480 Probes Master with Universal ProbeLibrary probes.

 

Contents

  • LightCycler® 480 Probes Master, 2x concentrated
  • Water, PCR Grade

The LightCycler® 480 Probes Master is designed for real-time PCR using the LightCycler® 480 Instrument in combination with suitable probes (e.g., hydrolysis probes, Universal ProbeLibrary probes, and others) and gene-specific primers. The kit is ideally suited for hot start PCR assays for gene quantification.

Hydrolysis probe assays use a single probe containing two labels: a fluorescent reporter dye and a quencher. While the probe is intact, the quencher is close to the reporter dye and suppresses the reporter fluorescence. When the probe is hybridized to the target sequence, the 5'-nuclease activity of the polymerase cleaves the hydrolysis probe, separating the reporter and quencher. With an increasing amount of target sequence during PCR, more probe is cleaved, and the fluorescence signal of the unquenched reporter dye increases.
The 2x concentrated master mix is optimized for a fixed MgCl2 concentration, which works with nearly all primer combinations. No adjustment in the MgCl2 concentration is needed to amplify different sequences; only template DNA, PCR primers, and hydrolysis probes (e.g., from the Universal ProbeLibrary) must be added.

Note: The LightCycler® 480 Probes Master has been specially optimized to use with hydrolysis probes (e.g.Universal ProbeLibrary probes). Other probe formats that can be adapted to use with the LightCycler® 480 Instrument include, for example, FRET (hybridization probes), Molecular Beacon, and Scorpions. Any fluorescent dye used in a LightCycler® 480 Instrument analysis must be compatible with the excitation and detection wavelengths provided with the LightCycler® 480 Instrument.
The LightCycler® 480 Probes Master can be used in conjunction with LightCycler® Uracil-DNA Glycosylase for carryover prevention during PCR. Furthermore, it can be used in two-step RT-PCR applications, for example, when used downstream of Transcriptor Reverse Transcriptase.


Quality

Function test: The LightCycler® 480 Probes Master is function tested using the LightCycler® 480 Instrument, according to the kit protocols.