FSTMPMMRO
Roche
FastStart TaqMan® Probe Master
sufficient for 100 reactions, sufficient for 500 reactions, sufficient for 2000 reactions, suitable for qPCR, suitable for RT-qPCR
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About This Item
UNSPSC Code:
41106300
NACRES:
NA.55
Recommended Products
usage
sufficient for 100 reactions
sufficient for 2000 reactions
sufficient for 500 reactions
Quality Level
feature
dNTPs included: no
hotstart
manufacturer/tradename
Roche
packaging
pkg of 100 x 50 μL reactions (04673409001)
pkg of 2000 x 50 μL reactions (04673433001)
pkg of 500 x 50 μL reactions (04673417001)
technique(s)
RT-qPCR: suitable
qPCR: suitable
input
purified DNA
detection method
probe-based
General description
FastStart TaqMan® Probe Master is a ready-to-use hot start reaction mix without ROX for quantitative polymerase chain reaction (qPCR) and reverse transcription (RT)-qPCR on real-time PCR systems other than the LightCycle® instruments. It is a 2x concentrated master mix that contains all the reagents (except primers, probe, and template). This master mix simplifies the preparation of reactions for DNA detection and analysis. It allows very sensitive detection and quantification of defined DNA sequences.
Hot start protocols have been also been shown to significantly improve the specificity 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 non-specifically 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).
This 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.
Combination of this master mix with our Transcriptor First Strand cDNA Synthesis Kit achieves excellent results in two-step qRT-PCR.
Hot start protocols have been also been shown to significantly improve the specificity 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 non-specifically 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).
This 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.
Combination of this master mix with our Transcriptor First Strand cDNA Synthesis Kit achieves excellent results in two-step qRT-PCR.
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).
Application
The FastStart TaqMan® Probe Master has been used in:
- quantitative, real-time DNA-detection assays
- qPCR and two-step reverse transcription (RT)-qPCR
- in the hydrolysis probe detection format
Features and Benefits
- Increase qPCR sensitivity and specificity:
- Use it with any probe-based assay:
- Amplify and detect a broad range of DNA or cDNA targets:
- Use with any real-time qPCR instrument other than the LightCycler® Instruments:
- Visualize amplification products on agarose gels.
- Use robotic pipetting stations to set up qPCR reactions:
- Prevent carryover contamination:
Components
FastStart TaqMan® Probe Master, 2x concentrated master mix that contains FastStart Taq DNA Polymerase, Reaction Buffer, and Nucleotides (dATP, dCTP, dGTP, dUTP).
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).
Other 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:
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.
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.
For life science research only. Not for use in diagnostic procedures.
Legal Information
LightCycler is a registered trademark of Roche
TaqMan is a registered trademark of Roche Molecular Systems, Inc.
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
does not flash
Flash Point(C)
does not flash
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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PCR master mix simplifies PCR/RT-PCR with components like DNA polymerase, dNTPs, MgCl2, and buffer, available commercially or DIY.
The purpose of Hot Start PCR is to inhibit the PCR reaction in order to reduce nonspecific amplification, prevent the formation of primer dimers, and increase product yields.
Related Content
Polymerase chain reaction (PCR) is a technique for amplifying nucleic acid molecules and is commonly used in many applications, including RT-PCR, hot start PCR, end point PCR and more.
RT-qPCR detects specific targets with applications in gene expression and pathogen detection.
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