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Radiant™ 2x RED Taq Mastermix , 200 Reactions, 5 x 1ml

$86.00

Radiant™ 2x RED Taq Mastermix , 200 Reactions, 5 x 1ml

$86.00
SKU:
RAD-200-5mL
Weight:
3.00 LBS
Shipping:
$50.00 (Fixed shipping cost)
Quantity:
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Product Description

Radiant™ Red 2X Taq Mastermix is a ready-to-use, robust PCR mix containing our highly purified Taq DNA Polymerase, reaction buffer, ultra-pure dNTPs, MgCl2 and PCR enhancers.

Radiant™ Red 2X Taq Mastermix is optimized for the sensitive DNA amplification of a wide range of DNA templates including complex mammalian genomic DNA and is ideal for screening, colony PCR, high-throughput PCR and genotyping of problematic GC-rich and AT-rich sequences. 

Radiant™ Red 2X Taq Mastermix is provided at 2× concentration and used at 1× concentration by adding template, primer, and H2O. 

  • Inert red dye will not inhibit PCR reaction & is used as a downstream Gel Loading dye
  • No change in PCR cycling conditions when in supplied Red format
  • New-generation PCR formulation including enhancers for maximum PCR efficiency, sensitivity and reaction speed.
  • Robust PCR performance across a wide range of DNA templates including genomic DNA and GC-rich and AT-rich sequences.
  • Supplied as a ready-to-use Red 2X Mastermix for maximum convenience and minimal liquid handling.
  • High-yields with amplicons up to 5 Kb with standard or fast cycling.

Storage

Radiant™ Red 2X Taq Mastermix is shipped on blue or dry ice and should be stored at -20˚C upon receipt. Excessive freeze/thawing should be avoided. When stored as specified, Radiant™ Red 2X Taq Mastermix is stable for 12 months from date of receipt. The Kit may also be stored at 4˚C for 1 month.

Important Considerations

Radiant™ Red 2X Taq Mastermix: The Red 2X Mastermix is comprised of Radiant™ Taq DNA Polymerase, Red color inert Gel Loading  Buffer, 2mM dNTPs, 6mM MgCl2, and PCR enhancers for maximum efficiency, sensitivity and success with difficult amplicons. We do not suggest the use of additional PCR enhancers.

Template: For complex genomic DNA, we suggest the use of 5ng  - 500ng per reaction; For cDNA or plasmid DNA, please use < 100ng per reaction.

Primers: Primers should have a predicted melting temperature of around 60°C, using default Primer 3 settings (http://frodo.wi.mit.edu/primer3/). The final primer concentration in the reaction should be between 0.2µM and 0.6µM.

Annealing: We recommend performing a temperature gradient to determine the optimal annealing temperature. Alternatively, we suggest a 55°C annealing temperature. Increase in 2°C increments if non-specific products are present. 

Extension: Optimal extension is achieved at 72°C. The optimal extension time is dependent on amplicon length and complexity. 15 seconds per kilobase(Kb) is recommended for amplification from eukaryotic genomic DNA or cDNA. For shorter amplicons, a 1 second extension is sufficient

Reaction setup

1. Prepare a PCR master mix based on following table:

Component         

50µl reaction                              

Final Concentration / Notes

Radiant™ Red 2X Taq Mastermix                           

25 µl                                              

1X

Forward Primer (10µM)                                       

2.0 µl                                             

400 nM

Reverse Primer (10µM)                                        

2.0 µl                                             

400 nM

Template DNA                                                 

<100ng cDNA                                     

<500ng genomic

Variable

PCR-grade water                                               

Up to 50 µl final volume

 

* For alternative total reaction volumes (eg. 25 µl), scale all components proportionally and maintain final concentrations

2.  PCR cycling:                

Cycles

Temperature & Time

Notes

1

95˚C                     2-5 minutes

Initial Denaturation

30-35

95˚C                     30 seconds

55˚C to 65˚C       30 seconds

72˚C                     60 seconds per Kb

Denaturation

Annealing

Extension

Quality Control

Radiant™ Red 2X Taq Mastermix is tested extensively for robust activity, processivity, efficiency, heat activation, sensitivity, absence of nuclease contamination and absence of nucleic acid contamination. Radiant™ Red 2X Taq Mastermix is manufactured under a comprehensive quality management system, following ISO 9001:2008 standards.

Limitations of Use

This product is intended for research purposes only and is not intended for any animal or human therapeutic use.

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