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Roche

FastStart Universal SYBR Green Master (Rox)

usage

sufficient for 200 reactions
sufficient for 2000 reactions

Quality Level

feature

dNTPs included: no
hotstart

packaging

pkg of 200 x 50 μL reactions (04913850001)
pkg of 2000 x 50 μL reactions (04913914001)

application(s)

RT-qPCR: suitable
qPCR: suitable

input

purified DNA

detection method

probe-based

General description

FastStart Universal SYBR® Green Master (Rox) is a ready-to-use hot start reaction mix for qPCR and RT-qPCR on all real-time PCR systems requiring normalization with ROX.

SYBR® Green I is a DNA double-strand-specific dye. During each phase of DNA synthesis, the SYBR® Green I dye, which is 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 (+15 to +25°C). Therefore, there is no elongation during the period when primers can non-specifically bind. 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

FastStart Universal SYBR® Green Master (Rox) can be used for the amplification and detection of any DNA or cDNA target, including those that are GC- or AT-rich.
Combine this master mix with Transcriptor First Strand cDNA Synthesis Kit (Roche) to achieve excellent results in two-step qRT-PCR.
FastStart Universal SYBR® Green Master (Rox) has been used in qRT-PCR and qPCR

Features and Benefits

  • Improve PCR sensitivity and specificity.
Minimize the formation of nonspecific amplification products.

  • Avoid over-estimation of qPCR results.
Eliminate nonspecific amplification products and primer-dimers that increase the amount of bound quantified SYBR Green I.
  • 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.

  • Save time and effort in qPCR preparation.
Eliminate the need to mix components, titrate MgCl2, or perform other time-consuming optimization steps.

  • Prevent false positives resulting from carryover contamination.
Use this dUTP-containing mix with Uracil-DNA Glycosylase to eliminate contaminating DNA carried over from previous PCR reactions.

Components

FastStart Universal SYBR Green Master (Rox), 2x concentrated master mix that contains FastStart Taq DNA Polymerase, Reaction Buffer, Nucleotides (dATP, dCTP, dGTP, dUTP), SYBR Green I, and a reference dye.

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

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

Legal Information

FastStart is a trademark of Roche
SYBR is a registered trademark of Life Technologies

Storage Class Code

12 - Non Combustible Liquids

WGK Germany

WGK 1

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Certificate of Analysis

Certificate of Origin

A novel deletion of IGF1 in a patient with idiopathic short stature provides insight Into IGF1 haploinsufficiency.
Batey L, et al.
The Journal of Clinical Endocrinology and Metabolism, 99(1), E153-E159 (2014)
Assessment of mTOR-dependent translational regulation of interferon stimulated genes.
Livingstone M, et al.
PLoS ONE, 10(7), e0133482-e0133482 (2015)
Lysine-specific demethylase 2B (KDM2B)-let-7-enhancer of zester homolog 2 (EZH2) pathway regulates cell cycle progression and senescence in primary cells.
Tzatsos A, et al.
The Journal of Biological Chemistry, 286(38), 33061-33069 (2011)
Regulation of vascular endothelial growth factor (VEGF) splicing from pro-angiogenic to anti-angiogenic isoforms a novel therapeutic strategy for angiogenesis.
Nowak D G, et al.
The Journal of Biological Chemistry, 285(8), 5532-5540 (2010)
The Kruppel-like factor 15 as a molecular link between myogenic factors and a chromosome 4q transcriptional enhancer implicated in facioscapulohumeral dystrophy.
Dmitriev P, et al.
The Journal of Biological Chemistry, 286(52), 44620-44631 (2011)

Articles

Hot Start PCR

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.

Hot Start PCR

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

PCR Applications

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 – Quantitative Reverse Transcription PCR

RT-qPCR, or quantitative reverse transcription PCR, combines the effects of reverse transcription and quantitative PCR or real-time PCR to amplify and detect specific targets. RT-qPCR has a variety of applications including quantifying gene expression levels, validating RNA interference (RNAi), and detecting pathogens such as viruses.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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