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FastStart TaqMan® Probe Master

Ready-to-use hot start reaction mix without ROX for qPCR and RT-qPCR on real-time PCR systems other than the LightCycler® instruments.

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

Product No. Pack Size List Price Your Price Quantity
04673409001
100 x 50 µL reactions $ 195.00

Product No. Pack Size List Price Your Price Quantity
04673417001
500 x 50 µL reactions $ 913.00

Product No. Pack Size List Price Your Price Quantity
04673433001
2000 x 50 µL reactions $ 3,374.00

The FastStart TaqMan® Probe Master is a ready-to-use reagent mix that simplifies the preparation of reactions for qPCR and two-step qRT-PCR. In combination with a real-time PCR instrument, suitable PCR primers, and a hydrolysis probe, FastStart TaqMan® Probe Master allows very sensitive detection and quantification of defined DNA sequences.
Do not use this product with the LightCycler® Instruments.
In principle, the FastStart TaqMan® Probe Master can be used for the amplification and detection of any DNA or cDNA target, including those that are GC- or AT-rich. However, you would need to adapt your detection protocol to the reaction conditions of the particular real-time PCR instrument used and design a specific hydrolysis probe and PCR primers for each target. See the Operator's Manual of your real-time PCR instrument for general recommendations.
Combine this master mix with our Transcriptor First Strand cDNA Synthesis Kit  to achieve excellent results in two-step qRT-PCR. Insist on the Transcriptor First Strand cDNA Synthesis Kit - designed and function-tested for qPCR, and efficient with all real-time PCR instruments.

 

  • Increase qPCR sensitivity and specificity:
    Produce lower cycle threshold (Ct) values with the hot start enzyme FastStart Taq DNA Polymerase and highest purity nucleotides included in the master mix.
  • Use it with any probe-based assay:
    Achieve sensitive, specific results in assays with the Universal ProbeLibrary Probes or any other hydrolysis probe.
  • Amplify and detect a broad range of DNA or cDNA targets:
    Amplify fragments up to 500 bp long, including those that are GC- or AT-rich.
  • Use with any real-time qPCR instrument other than the LightCycler® Instruments:
    Choose from two formulations - one that contains the ROX reference dye and one without ROX.
  • Visualize amplification products on agarose gels.
  • Use robotic pipetting stations to set up qPCR reactions:
    Benefit from a master mix that is stable at room temperature during extended reaction setup times.
  • Prevent carryover contamination:
    The mix contains dUTP, so that it may be used with Uracil-DNA Glycosylase to prevent false positives arising from carryover contamination (i.e., contamination with amplified DNA).

Contents

FastStart TaqMan® Probe Master, 2x concentrated master mix that contains FastStart Taq DNA Polymerase, Reaction Buffer, and Nucleotides (dATP, dCTP, dGTP, dUTP).

The FastStart TaqMan® Probe Master is a ready-to-use, 2x concentrated master mix that contains all the reagents (except primers, probe, and template) needed for running quantitative, real-time DNA-detection assays, including qPCR and two-step qRT-PCR, in the hydrolysis probe detection format. Use the FastStart TaqMan® Probe Master with any real-time qPCR instrument other than the LightCycler® Instruments.



Figure 1: Sensitive detection of an RNA template with the Transcriptor First Strand cDNA Synthesis Kit and the FastStart TaqMan® Probe Master. One hundred nanograms of total RNA (control RNA from a control kit of Supplier A) was reverse transcribed with the Transcriptor First Strand cDNA Synthesis Kit (+55°C, 30 minutes, 20 µL final volume). The cDNA primers were specific for the GAPDH gene. A portion (2 µL) of the cDNA product was used as template in a qPCR. The PCR primers and a hydrolysis probe were also specific for the GAPDH gene. This reaction was performed twice, once with the FastStart TaqMan® Probe Master and once with a kit from another supplier. Both qPCRs were run on an Applied Biosystems 7500 Real-Time PCR System. All reactions were performed in duplicate; the ROX reference dye was included in every reaction.
Result: The reactions with the Transcriptor First Strand cDNA Synthesis Kit and the FastStart TaqMan® Probe Master produced the best sensitivity.

Hot start protocols have been shown to significantly improve the specificity, sensitivity, and yield of PCR. Heat-labile blocking groups on some of the amino acid residues of FastStart Taq DNA Polymerase make the modified enzyme inactive at room temperature. Therefore, there is no elongation during the period when primers can nonspecifically bind. The FastStart Taq DNA Polymerase is activated by removing the blocking groups at a high temperature (i.e., a pre-incubation step at +95°C).
 

Quality

Function test: Each lot is tested for performance in qPCR using three templates: a GC-rich template, an AT-rich template, and a long template (approximately 440 bp).


Notes

qPCR targets
In principle, the FastStart TaqMan® Probe Master can be used for the amplification and detection of any DNA or cDNA target, including those that are GC- or AT-rich. However, for each target, you would need:

  • to adapt your detection protocol to the reaction conditions of your real-time PCR instrument, and
  • design specific PCR primers and hydrolysis probes for the target.

See the Operator's Manual of your real-time PCR instrument for general recommendations.
Two forms available
The master mix is available in two forms – one that contains the ROX reference dye and one without ROX.
Preventing carryover contamination
This master contains dUTP, which will be incorporated into PCR products to help prevent false positives resulting from carryover contamination. In subsequent PCRs, you can add Uracil-DNA Glycosylase to degrade any uracil-containing carryover contaminants (amplification products from previous PCRs).
qRT-PCR
Combine this master mix with our Transcriptor First Strand cDNA Synthesis Kit for two-step qRT-PCR. This kit gives excellent results and works efficiently with all real-time PCR instruments. (See "Typical Experiment" above.)

Product No: 04673409001
Product No: 04673417001
Product No: 04673433001

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