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- Radiant 2x RED Taq Mix, 1000 Reactions, 25 x 1ml
- Home
- Molecular Biology
- Radiant 2x RED Taq Mix, 1000 Reactions, 25 x 1ml
Radiant 2x RED Taq Mix, 1000 Reactions, 25 x 1ml
- SKU:
- RAD-1000-1mL
- Weight:
- 3.00 LBS
- Shipping:
- Calculated at Checkout
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.