GreenMasterMix (2X) No ROX for qPCR
Order number: M3023.0100
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GreenMasterMix without 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 Mastermix is ready-to-use and contains optimised amounts of all ingredients.
Advantages of the chemically modified SuperHot Taq polymerase used in the Genaxxon GreenMasterMix:
- 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 Green 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
- intercalating green fluorescent dye
- 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: BioRad CFX96 Touch™, CFX384 Touch™, CFX Connect™, DNA Engine Opticon® 2, Chromo4™, iCycler iQ™ and My iQ™ , Roche LightCycler® 480, LightCycler® 1536, LightCycler® Nano, LightCycler® 96 and QuantStudio™ instruments, Thermo Scientific™ PikoReal™, Cepheid SmartCycler®, Bio Molecular Systems Mic qPCR cycler, Qiagen Rotor Gene Q, Rotor Gene 6000, MyGo Mini and MyGo Pro.
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:
Specifications:
This realtime PCR master mix is a ready-to-use 2-time master mix that contains no ROX™
Suited for FAST PCR with MIC Cycler
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 GreenMasterMix (2X) No 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 (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.
Referenzen..
Hier finden Sie Artikel und Literaturzitate, in denen die Autoren auf die hohe Qualität dieses Genaxxonprodukts vertrauen.
Listed below are articles and references, in which the authors trust in the high quality of this Genaxxon product.
Quelle/Source: NCBI PubMed >
Keratinocyte-derived IκBζ drives psoriasis and associated systemic inflammation
Sebastian Lorscheid, Anne Müller, Jessica Löffler, Claudia Resch, Philip Bucher, Florian C. Kurschus, Ari Waisman, Knut Schäkel, Stephan Hailfinger, Klaus Schulze-Osthoff, Daniela Kramer
JCI Insight. 2019 Nov 14; 4(22): e130835. Published online 2019 Nov 14. doi: 10.1172/jci.insight.130835
PMCID: PMC6948851
Long Noncoding RNA SSR42 Controls Staphylococcus aureus Alpha-Toxin Transcription in Response to Environmental Stimuli
Jessica Horn, Maximilian Klepsch, Michelle Manger, Christiane Wolz, Thomas Rudel, Martin Fraunholz
J Bacteriol. 2018 Nov 15; 200(22): e00252-18. Prepublished online 2018 Aug 27. Published online 2018 Oct 23. doi: 10.1128/JB.00252-18
PMCID: PMC6199474
MiR-744-5p inducing cell death by directly targeting HNRNPC and NFIX in ovarian cancer cells
Michael Kleemann, Helga Schneider, Kristian Unger, Philip Sander, E. Marion Schneider, Pamela Fischer-Posovszky, René Handrick, Kerstin Otte
Sci Rep. 2018; 8: 9020. Published online 2018 Jun 13. doi: 10.1038/s41598-018-27438-6
PMCID: PMC5998049
miR-217-5p induces apoptosis by directly targeting PRKCI, BAG3, ITGAV and MAPK1 in colorectal cancer cells
Marion Flum, Michael Kleemann, Helga Schneider, Benjamin Weis, Simon Fischer, René Handrick, Kerstin Otte
J Cell Commun Signal. 2018 Jun; 12(2): 451–466. Published online 2017 Sep 14. doi: 10.1007/s12079-017-0410-x
PMCID: PMC5910322
