ProbeMasterMix (2X) Low ROX for qPCR
Order number: M3031.0100
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ProbeMasterMix low ROX optimised for realtime PCR assays in block systems that contains all components to perform quantitative PCR with the exception of primer and template DNA. This 2X master mix is ready-to-use and contains optimised amounts of all ingredients.
Multiplex PCR: Applications at Genaxxon and at customers site have shown that the qPCR Probe Mastermix can be used for the simultaneous detection of up to four DNA targets in the same PCR reaction. For further details please refer to the product manual or contact us: info@genaxxon.com.
Advantages of the Genaxxon ProbeMasterMix:
- Hotstart technology enables setup of PCR mixture at room temperatur
- 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 SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity:
1. The chemical modification 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 without ROX contains all the necessary components in an optimized composition to carry out quantitative PCR.
- chemically modified Taq DNA Polymerase
- dATP, dCTP, dGTP, dTTP
- 50nM ROX
- optimized reaction buffer
- 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)
Stability at +2°C to +8°C (refrigerator): minimum of 8 months.
This Mastermix is specially suited for the following instruments: Applied Biosystems® 7500, 7500 Fast and ViiA™ 7, QuantStudio™ instruments, Agilent Mx3000P™, Mx3005P™, Mx4000™ and AriaMx.
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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High Sensitivity and Specifity, Optimised buffer system, Reproducibility over a wide range of template amounts, Broad dynamic range. Aliquot size: 1.25mL. This master mix is stable for at least 8 months at +2°C to +8°C.
Technical Data:
2-time ready-to-use realtime PCR mastermix with 50nM ROX™ as reference dye.
Aliquot size: 1.25mL. This master mix is stable for at least 8 months at +2°C to +8°C.
application:
PCR Primer extension Multiplex PCR Low-copy targets PCR Real-time PCR Quantitative PCRUnit 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 / Classification
eclass-Nr: 34-16-04-90Dokumente - Protokolle - Downloads
Here you will find information and further literature on ProbeMasterMix (2X) Low 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: info@genaxxon.com or phone: +49 731 3608 123.
Dokumente - Protokolle - Downloads
Fast PCR:
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 on Mx3005P
GreenMasterMix Low ROXTM and ProbeMasterMix Low ROXTM were tested on Mx3005P (Agilent Technologies) using fast cycling real-time 2-step protocols; Fast Pthr Green Mx and Fast Pthr Probe Mx, respectively.
qPCR Setup: For GreenMasterMix Low ROXTM and ProbeMasterMix Low ROXTM, primers targeting 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. 3 and 4. The PCR reaction mix was run on MX3005P (Requiring Low ROX). Instrument settings, run times and quality criteria for the applied fast protocols; Fast Pthr Green Mx and Fast Pthr Probe Mx, were compared to the standard protocol; Standard Pthr Mx. See table 2.
Results: The standard Fast Pthr Probe Mx protocol has a run time of 83.4 minutes. Fast Pthr Green Mx and Fast Pthr Probe Mx protocols, resulted in run times of 50.1 and 46.8 minutes, respectively. All protocol times are inclusive ramping time and a15 minutes hot start. The determined criteria, as shown in table 2, for Fast Pthr Green Mx and Fast Pthr Probe Mx protocols were acceptable. Furthermore, the melt curve analysis using GreenMasterMix Low ROXTM with Fast Pthr Green Mx protocol, confirmed high specificity.
Figure 3. Amplification plot, standard curve and melt curve for GreenMasterMix run on Mx3005P using Fast Pthr Green Mx protocol. Primers targeting 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.
Figure 4. Amplification plot and standard curve for ProbeMasterMix run on Mx3005P using Fast Pthr Probe Mx protocol. Primers targeting Pthr (75 bp) and 4 concentrations of gDNA (40 ng, 20 ng, 10 ng and 5 ng) were used.
