FSSGMMRO
Roche
FastStart SYBR Green Master
sufficient for 500 reactions, sufficient for 5000 reactions, suitable for qPCR, suitable for RT-qPCR
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About This Item
UNSPSC Code:
41106300
Recommended Products
usage
sufficient for 500 reactions
sufficient for 5000 reactions
Quality Level
feature
dNTPs included: no
hotstart
manufacturer/tradename
Roche
packaging
pkg of 500 x 20 μL reactions (04673484001)
pkg of 5000 x 20 μL reactions (04673492001)
technique(s)
RT-qPCR: suitable
qPCR: suitable
input
purified DNA
detection method
probe-based
Related Categories
General description
FastStart SYBR Green Master; Instructions For Use
FastStart™ SYBR® Green 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 LightCycler® instruments. This master mix simplifies the preparation of reactions for DNA detection and analysis. 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.
SYBR® Green I is a DNA double-strand-specific dye. During each phase of DNA synthesis, the SYBR® Green I dye, included in the reaction mix, binds to the amplified PCR products. The amplicon can be detected by its fluorescence.
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 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).
The FastStart™ SYBR® Green 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 specific PCR primers for each target.
Combine this master mix with our Transcriptor First Strand cDNA Synthesis Kit to achieve excellent results in two-step qRT-PCR.
FastStart™ SYBR® Green 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 LightCycler® instruments. This master mix simplifies the preparation of reactions for DNA detection and analysis. 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.
SYBR® Green I is a DNA double-strand-specific dye. During each phase of DNA synthesis, the SYBR® Green I dye, included in the reaction mix, binds to the amplified PCR products. The amplicon can be detected by its fluorescence.
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 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).
The FastStart™ SYBR® Green 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 specific PCR primers for each target.
Combine this master mix with our Transcriptor First Strand cDNA Synthesis Kit to achieve excellent results in two-step qRT-PCR.
Application
The FastStart™ SYBR® Green Master has been used in qPCR and two-step qRT-PC in the SYBR® Green I detection format. It is also used:
Use the FastStart™ SYBR® Green Master with any real-time qPCR instrument other than the LightCycler® Instruments.
- in quantitative polymerase chain reaction (qPCR) for adeno-associated virus (AAV) titre quantification
- in qPCR to determine the expression level of different Col6a1-3 genes relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH)
- in quantitative real-time polymerase chain reaction (qRT-PCR) of G protein-coupled estrogen receptor-1 (GPER) mRNA
- in qRT-PCR for gene expression studies
Use the FastStart™ SYBR® Green Master with any real-time qPCR instrument other than the LightCycler® Instruments.
Features and Benefits
- Improve PCR sensitivity and specificity:
- Avoid over-estimation of qPCR results:
- Amplify and detect a broad range of DNA or cDNA targets:
- Use any real-time PCR instrument other than the LightCycler® Instruments:
- Save time and effort in qPCR preparation:
- Prevent false positives resulting from carryover contamination:
Components
FastStart SYBR Green Master, 2x concentrated master mix that contains FastStart Taq DNA Polymerase, Reaction Buffer, Nucleotides (dATP, dCTP, dGTP, dUTP), and SYBR Green I.
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 (about 440 bp).
Other Notes
qPCR targets
In principle, the FastStart SYBR Green 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 SYBR Green 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 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
FastStart is a trademark of Roche
LightCycler is a registered trademark of Roche
SYBR is a registered trademark of Life Technologies
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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Articles
A PCR master mix is a batch of PCR or RT-PCR reagents that can be divided among many PCR reaction tubes. It usually includes DNA polymerase, dNTPs, MgCl2 and buffer. Make your own master mix or choose a commercial one.
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.
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