HotRox Master Mix 2x
| Catalog # | Pack size | Price, Euro | SHOP |
| 119550 | HotRox Master Mix 2x (Final ROX conc. 500 nM; 100 rxn) (2x1,25ml) | 75,00 | Add to cart |
| 119505 | HotRox Master Mix 2x (Final ROX conc. 50 nM; 100 rxn) (2x1,25ml) | 72,00 | Add to cart |
Research use only
Datasheet: HotRox Master Mix 2x
Description:
2 x HotRox Master Mix is an optimized ready-to-use PCR mixture of Taq DNA Polymerase, antibodies to Taq DNA Polymerase, PCR buffer, MgCl2 and dNTPs. 2x PCR Master Mix contains all components for PCR, except DNA template and primers. ROX Reference dye is included in the HotRox Master Mix to normalize the fluorescent signal on instruments that are compartible with this option. The mixture was shown to be effective for Real Time PCR.
2x HotRox Master Mix Composition
- Taq DNA Polymerase (recombinant) in reaction buffer: 0.1 unit/µl
- Antibodies to Taq DNA Polymerase, concentration adjusted for the effective inhibition of DNA polymerase activity at 37oC
- 32 mM (NH4)2SO4
- 130 mM TrisHCl, pH 8.8 at 25oC
- 0.02% Tween-20
- 3 mM MgCl2
- dNTPs (dATP, dCTP, dGTP, dTTP): 0.4 mM of each
Instrument | Final ROX Conc. |
| ABI 7000, 7300, 7700, 7900HT | 500 nM |
| ABI 7500, Stratagene Mx3005P, Mx4000 | 50nM |
Pack size:
2x 1,25 ml - sufficient for 100 HotStart PCR reactions in 50 µl reaction volume
Performance and purity tests :
Tested for the absence of endodeoxyribonucleases and exodeoxyribonucleases. The 2x HotRox Master Mix is tested in the amplification of a single-copy gene of mouse genomic DNA.
Endodeoxyribonuclease Assay:
No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 25 µl of 2x HotRox Master Mix with 1 µg of pUC19 DNA in 50 µl for 4 hours neither at 37°C nor at 70oC.
Application:
- PCR
- Primer extension
- Multiplex PCR
- Low-copy targets PCR
- Real-time PCR
Associated activities:
Endonuclease and exonuclease activities were not detectable after 2 hours incubation of the mixture with 0.22 mg of EcoR I digested lambda DNA at 72oC in the presence of 15-20 units of enzyme in HotRox Master MIX
Storage conditions:
Store HotRox Master MIX frozen at -20°C and protect from light.
Protocol for PCR with SuperHot PCR Master Mix
Due to the inhibition of polymerase activity at room temperature by Anti Taq DNA polymerase antibodies all reactions may be settled-up at room temperature, it will not result in increase of unspecific product or primer-dimers formation.
Add in a thin walled PCR tube:
| 50 µl reaction volume | 25 µl reaction volume | |||
| component | volume | final conc. | volume | final conc. |
| 2x PCR Master Mix | 25 µl | 1x | 12.5 µl | 1x |
| Forward Primer | variable | 0.1-1 µl | variable | 0.1-1 µl |
| Reverse Primer | variable | 0.1-1 µl | variable | 0.1-1 µl |
| Template DNA | variable | 10 pg-1µg | variable | 10 pg-1 µg |
| Sterile Deionized Water | up to 50 µl | - | up to 25 µl | - |
- Gently vortex the sample and briefly centrifuge to collect all drops to the bottom of the tube.
- Overlay the sample with mineral oil or add an appropriate amount of wax if the thermal cycler is not equipped with a heated lid.
- Place the samples in a thermocycler and start the optimal PCR program.
