Order number: M3155.1000Shipping: shipped on wet ice, store at -20°C
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Delivery time: 3- 8 working days
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ProbeMasterMix FAST with 500nM ROX optimised for fast realtime PCR assays in block systems. This master mix is ready-to-use and contains all components for a successful and reliable quantitative PCR with the exception of primer and template DNA.
Advantages of the Hotstart Taq polymerase used in the Genaxxon ProbeMasterMix:
- Hotstart technology enables setup of PCR mixture at room temperature.
- short initial denaturation of not more than 2 minutes.
- optimized for 2-step PCR protocols.
- No pipetting on ice necessary anymore
- No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days.
- Amplification of GC-rich templates
- High yields
- No Primer dimers
The Genaxxon Hotstart Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity:
1. The hotstart formulation inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and
2. High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results.
The Probe qPCR master mix with 500nM ROX contains all the necessary components in an optimized composition to carry out quantitative PCR.
- hotstart Taq DNA Polymerase
- dATP, dCTP, dGTP, dTTP
- optimized reaction buffer
- 500nM ROX as internal reference
- stabilizers and enhancers to enable even amplification of low copy number targets
- the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote)
This Mastermix is specially suited for the following instruments: Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900 HT, Eppendorf Realplex4, StepOne™ and StepOnePlus™.
Other realtime PCR master mixes from Genaxxon are: M3023 - GreenMastermix No ROX >, M3045 - ProbeMastermix No ROX >, M3011 - GreenMastermix Low ROX >, M3031 - ProbeMastermix Low ROX >, M3052 - GreenMastermix High ROX >, M3010 - ProbeMastermix High ROX >.
Test sample available! The test sample price will be refunded on the first official order of the product.
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- Further products by Genaxxon bioscience
- FAST start system. Almost no initial denaturation needed! - High Sensitivity and Specifity - Optimised buffer system - Reproducibility over a wide range of template amounts - Broad dynamic range. - Aliquot size: 1.25mL.
This ProbeMastermix is a ready-to-use 2-time master mix that contains no ROX™
short initial denaturation times
optimized for 2-step qPCR
Aliquot size: 1.25mL.
application:PCR Primer extension Multiplex PCR Low-copy targets PCR Real-time PCR Quantitative PCR
Unit Definition:One unit is defined as the amount of enzyme which will convert 10 nmoles of dNTPs to an acid-insoluble form in 30 min at 72°C under the assay conditions (25 mM TAPS (tris-(hydroxymethyl)-methyl-amino-propanesulfonic acid, sodium salt) pH 9.3 (25°C); 50 mM KCl; 2 mM MgCl2; 1 mM b-mercaptoethanol; and activated calf thymus DNA as substrate.
Sicherheits Hinweise / Safety
Klassifizierungen / Classificationeclass-Nr: 34-16-04-90
Dokumente - Protokolle - Downloads
Here you will find information and further literature on ProbeMasterMix FAST with 500nM ROX for qPCR. For further documents (certificates with additional lot numbers, safety data sheets in other languages, further product information) please contact Genaxxon biosience at: email@example.com or phone: +49 731 3608 123.
Dokumente - Protokolle - Downloads
When performing fast real-time PCR, it is of high importance to consider primer design and target size. For efficient amplification during fast cycling conditions, target size between 70 bp and 200 bp is recommended. The shorter the target length, the faster the total run time. For other targets and other primer sets, the annealing step has to be determined experimentally.
Please use the following protocol for inspiration as you need to optimize instruments settings according to yourr target size and applied instruments.
Fast real-time PCR results using Genaxxon Master Mixes (without ROX) on LightCycler® 96
GreenMasterMix (2X) without ROX or ProbeMasterMix (2X) without ROX was tested on LighCycler96 (Roche) using fast cycling real-time 2-step protocols; Fast PAH12 Green LC and Fast Pthr Probe LC, respectively.
qPCR Setup: GreenMasterMix (2X) (without ROX) or ProbeMasterMix (2X) (without ROX), primers targeting either PAH12 (203 bp) or Pthr (75 bp) and 4 concentrations of gDNA (40 ng, 20 ng, 10 ng and 5 ng) were used. All DNA concentrations were tested in quadruple replicates. See fig. 1 and 2. The PCR reaction mix was run on LightCycler® 96 (No requirements for ROX). Instrument settings, run times and quality criteria for the applied fast protocols; Fast PAH12 Green LC and Fast Pthr Probe LC, were compared to the standard protocol; Standard Pthr LC. See table 1.
Results: The standard Fast Pthr Probe LC protocol has a run time of 80.8 minutes. Fast PAH12 Green LC and Fast Pthr Probe LC protocols, resulted in run times of 47.5 and 40.8 minutes, respectively. All protocol times are inclusive ramping time and a15 minutes hot start. The determined criteria, as shown in table 1, for Fast PAH12 Green LC and Fast Pthr Probe LC protocols were satisfying. Furthermore, the melt curve analysis using Green 2x Master Mix without ROX the Fast PAH12 Green LC protocol confirmed high specificity.