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Red MasterMix (2x) - PCR MasterMix with red dye

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  • High-quality products
  • Customized solutions
  • Personal contact
  • Fast service
  • Competent technical support +49(0)731-3608-123


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Product number: M3029.2500
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Shipment: on wet ice. Stored at -20°C. For laboratory usage only!
Product information "Red MasterMix (2x) - PCR MasterMix with red dye"

Red MasterMix (2x) - PCR MasterMix with red dye to help visualize pipetting and mixing steps. Electrophoresis can be performed immediately after PCR without the need of a gel loading buffer. This makes this MasterMix time efficient, cost efficient and a reliable choice for the best PCR results with high efficiency.  One tube, one pipetting step.

Red MasterMix (2x) is delivered to you as a premixed, ready-to-use solution containing Taq DNA Polymerase, dNTPs, MgCl2 and reaction buffers at optimal concentrations. Additionally the Red MasterMix (2x) contains an additive and a red dye for proceeding with electrophoresis after PCR without adding loading buffer. Just add your primers and template DNA.
This PCR MasterMix (2x) has been optimized for use in routine PCR amplification of DNA templates in the range of 0.2-4kb. The special formulation of our PCR RedMasterMix (2x) can withstand repeated freezing/thawing without compromising yields, sensitivity or results, when used as directed.

The PCR MasterMix with red dye is efficient (no left over of reagents), scalable from 10µL to 50µL and stable for at least 24 months.

Our RedMastermix can also be used for Sanger sequencing. Just dilute the PCR reaction 1:8 after PCR, or purify using spin columns and then apply.

Read now in our blog how Genaxxon's Red MasterMix can also simplify your laboratory work.

Our Agaroses > and DNA Ladders > are ideally suited for subsequent electrophoresis of PCR products.

FAQs
No search results.
No, the included red dye does not interfere with the PCR performance. If necessary, the red dye can be removed, e.g., by spin column purification.
Yes, you can use your own cycling conditions. Please note that the cycling conditions in our protocol are only guidelines for PCR amplification. Optimal reaction conditions such as incubation times, temperatures, and amount of template DNA may vary and must be determined individually.
You need approximately 10E4 copies of target DNA to detect product in 25-30 PCR cycles.
The Red MasterMix provides a final concentration of 1.5mM MgCl2 which will give satisfactory results in most cases. However, the ideal concentration depends on the template, buffer, DNA and dNTPs, as each has the potential to chelate magnesium. If the Mg2+ concentration is too low, no PCR products will be seen. In these cases it may be necessary to add additional MgCl2.

Specifications:
2-time ready-to-use master mix for PCR including a red dye for better visualization of pipetting and as gel loading dye. dNTPs, buffer and DNA polymerase already included.

Convenient Aliquot size: 1.25mL. This master mix is stable for at least 8 months at +2°C to +8°C.


Applikation / Application:
For reliable PCR from mouse tails. For Sanger sequencing reactions.

Einheiten / Units:
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.

Quelle / Source:
synthetic


Sicherheits Hinweise / Safety


Klassifizierungen / Classification

eclass-Nr: 32-16-05-02
Documents - Protocols - Downloads :
Here you will find information and further literature. 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.


Documents:

Safety Data Sheet
Protocols
Manuals
General Data 1
Category List
Listed below are articles and references, in which the authors trust in the high quality of this Genaxxon product.
Source: NCBI PubMed

References

Listed below are articles and references in which the authors trust in the high quality of this Genaxxon product. 

Source: NCBI PubMed >

Widespread co-endemicity of Trypanosoma species infecting cattle in the Sudano-Sahelian and Guinea Savannah zones of Cameroon
Archile Paguem, Babette Abanda, Dieudonné Ndjonka, Judith Sophie Weber, Sen Claudine Henriette Ngomtcho, Kingsley Tanyi Manchang, Mamoudou Adoulmoumini, Albert Eisenbarth, Alfons Renz, Sørge Kelm, Mbunkah Daniel Achukwi
BMC Vet Res. 2019; 15: 344. Published online 2019 Oct 16. doi: 10.1186/s12917-019-2111-6
PMCID: PMC6796345

Spatiotemporal Role of Transforming Growth Factor Beta 2 in Developing and Mature Mouse Hindbrain Serotonergic Neurons
Enaam Chleilat, Robert Mallmann, Rainer Spanagel, Norbert Klugbauer, Kerstin Krieglstein, Eleni Roussa
Front Cell Neurosci. 2019; 13: 427. Published online 2019 Sep 20. doi: 10.3389/fncel.2019.00427
PMCID: PMC6763588

Molecular identification and prevalence of tick-borne pathogens in zebu and taurine cattle in North Cameroon
Babette Abanda, Archile Paguem, Mamoudou Abdoulmoumini, Manchang Tanyi Kingsley, Alfons Renz, Albert Eisenbarth
Parasit Vectors. 2019; 12: 448. Published online 2019 Sep 11. doi: 10.1186/s13071-019-3699-x
PMCID: PMC6737592

Impact of Alternatively Polyadenylated Isoforms of ETHYLENE RESPONSE FACTOR4 with Activator and Repressor Function on Senescence in Arabidopsis thaliana L.
Lena Riester, Siliya Köster-Hofmann, Jasmin Doll, Kenneth W. Berendzen, Ulrike Zentgraf
Genes (Basel) 2019 Feb; 10(2): 91. Published online 2019 Jan 28. doi: 10.3390/genes10020091
PMCID: PMC6409740

Recruitment of Cytosolic J-Proteins by TOM Receptors Promotes Mitochondrial Protein Biogenesis
Łukasz Opaliński, Jiyao Song, Chantal Priesnitz, Lena-Sophie Wenz, Silke Oeljeklaus, Bettina Warscheid, Nikolaus Pfanner, Thomas Becker
Cell Rep. 2018 Nov 20; 25(8): 2036–2043.e5. Published online 2018 Nov 20. doi: 10.1016/j.celrep.2018.10.083
PMCID: PMC6280124

Membrane protein insertion through a mitochondrial β-barrel gate

Alexandra I.C. Höhr, Caroline Lindau, Christophe Wirth, Jian Qiu, David A. Stroud, Stephan Kutik, Bernard Guiard, Carola Hunte, Thomas Becker, Nikolaus Pfanner, Nils Wiedemann

Science. Author manuscript; available in PMC 2018 Jul 19.

Published in final edited form as: Science. 2018 Jan 19; 359(6373): eaah6834. doi: 10.1126/science.aah6834

PMCID: PMC5959003

 

Nimodipine fosters remyelination in a mouse model of multiple sclerosis and induces microglia-specific apoptosis

Andrea Schampel, Oleg Volovitch, Tobias Koeniger, Claus-Jürgen Scholz, Stefanie Jörg, Ralf A. Linker, Erhard Wischmeyer, Marie Wunsch, Johannes W. Hell, Süleyman Ergün, Stefanie Kuerten

Proc Natl Acad Sci U S A. 2017 Apr 18; 114(16): E3295–E3304. Published online 2017 Apr 5. doi: 10.1073/pnas.1620052114

PMCID: PMC5402421

 

Interleukin-4 Protects Dopaminergic Neurons In vitro but Is Dispensable for MPTP-Induced Neurodegeneration In vivo

Laura Hühner, Jennifer Rilka, Ralf Gilsbach, Xiaolai Zhou, Venissa Machado, Björn Spittau

Front Mol Neurosci. 2017; 10: 62.  Published online 2017 Mar 9. doi: 10.3389/fnmol.2017.00062

PMCID: PMC5343015

Asexual Recombinants of Plasmopara halstedii Pathotypes from Dual Infection of Sunflower

Otmar Spring, Reinhard Zipper

PLoS One. 2016; 11(12): e0167015.  Published online 2016 Dec 1. doi: 10.1371/journal.pone.0167015

PMCID: PMC5132302

 

Molecular identification and prevalence of tick-borne pathogens in zebu and taurine cattle in North Cameroon

Babette Abanda, Archile Paguem, Mamoudou Abdoulmoumini, Manchang Tanyi Kingsley, Alfons Renz, Albert Eisenbarth

Parasit Vectors. 2019; 12: 448. Published online 2019 Sep 11. doi: 10.1186/s13071-019-3699-x

PMCID: PMC6737592


For every template/primer pair, the optimal reaction conditions have to be evaluated empirically, changing the primer/template
ratio, the ionic strength (with MgSO4) and the cycle parameters (time and temperatures).

DNA Template

Amplification of templates with high GC content, strong secondary structure, low concentrations, or which produce products greater than 5 kb, may require adaptation of the following parameters:

  • The Genaxxon RedMastermix is particularly well suited for less clean mouse tail DNA
  • Approximately 10E4 copies of target DNA are required to detect product in 25-30 PCR cycles.
  • Use 1pg–1ng of plasmid or viral templates.
  • Use 1ng–1µg of genomic templates.
  • Higher DNA concentrations decrease amplicon specificity (i.e., extra bands are more likely), particularly when a large number of cycles are employed.
  • Use the higher DNA concentrations when fewer cycles are desired (e.g. to increase fidelity).

Primers

  • Generally 20-30 nucleotides in length.
  • Ideal GC content is 40-60%.
  • Space GC residues evenly within the primer.
  • Primer pairs should have Tms within 5°C of each other.
  • Avoid secondary structure (i.e., hairpins) within each primer and potential dimerization between the primers present.
  • When engineering sites into the end of primers, 4-6 extra bases should be added 5´ to the site.
  • Final concentration should be 0.05-1 µM, typically 0.1-0.5 µM of each primer.
  • Higher concentrations may increase secondary priming and create spurious amplification products.

Magnesium Concentration

  • 1.5-2.0 mM is optimal for Taq DNA Polymerase, but the ideal concentration depends on template, buffer, DNA and dNTPs (each has the potential to chelate magnesium).
  • If [Mg2+] is too low, no PCR product will be seen.
  • If [Mg2+] is too high, undesired PCR products may be seen.

Deoxyribonucleotide triphosphates (dNTPs) >

  • Typical concentration is 200 µM of each dNTP.
  • 50-100 µM enhances fidelity of polymerization, but reduces yields.
  • Higher concentrations increase yields particularly in long PCR, but can reduce fidelity.

DNA Polymerase

  • The choice of the correct polymerase depends among other things on the purpose as well as on the template used (standard PCR: Taq DNA Polymerase S > with high accuracy, Taq Polymerase E > with high yield; master mixes: Standard PCR MasterMix > or RedMasterMix > with red dye).
  • For multiplex PCR, there exist special Multiplex MasterMixes >.
  • Hot start applications are recommended to increase specificity or if you use difficult templates. For this purpose there exist special Hot Start Polymerases >.
  • For PCRs for the purpose of cloning or other procedures requiring a low error rate, there are thermostable high fidelity proofing polymerases such as Pfunds >, ExactRun >, ReproFast >, or ReproHot (KOD) Proofreading Polymerase >. These enzymes make far fewer errors during amplification and increase the chances of an amplicon without mistakes.
  •  For genotyping and other applications where a high discrimination rate is required, there is a new highly selective DNA polymerase, SNP Pol DNA Polymerase >. It specifically distinguishes mismatched primer-template complexes and ony produces specific amplicons in case of perfectly matched primer pairs.
  • For the real-time quantitative PCR > there exist highly specialized qPCR master mixes. Hereby, the choice of the correct master mix depends on the qPCR device you are using. It is important to determine whether and how much ROX should be contained in the qPCR master mix and whether it should be a qPCR green master mix with green fluorescent dye > or a qPCR probe  master mix > without a fluorescent dye. A new lyophilized qPCR probe master mix in beads format, which is stable at room temperature, is available from Genaxxon (LyoBalls >).
  • The amount of DNA polymerase used in the PCR reaction can significantly influence the PCR result (use 1.25-1.5 units Taq Polymerase > for a 50μL volume).

General Guidelines

Annealing Temperature and Duration

  • Match the Tms within 5°C of each other.
  • Typical annealing temperatures are 5°C below the lowest primer's Tm and often fall in the range of 50-60°C.
  • Test higher annealing temperatures if spurious amplification products are observed.
  • Typical annealing times are 15-30 seconds.

Extension Time

  • Extensions are normally performed at 68°C.
  • In general, use extension times of one minute per 1000 base (1 kb) pairs (e.g. 3 minutes for a 3 kb product). 
  • For products less than 1 kb, use 45-60 seconds.
  • Products greater than 3 kb, or reactions using more than 30 cycles, may require longer extensions.

The amplification of templates with high GC content, strong secondary structures, low concentrations or amplicons more than 5 kb often require optimization of the PCR conditions. Typically, 15-30 seconds of denaturation should be performed at 95 ° C during PCR. For every template/primer pair the optimal reaction conditions have to be evaluated empirically, changing the primer/template ratio, the ionic strength ( with MgSO4) and the cycle parameters (time and temperatures).