dU-HOT Mastermix
| Catalog # | Pack size | Price, Euro | SHOP |
| 119202 | 100 rcs (2x1,25ml) | 65,00 | Add to cart |
| 119210 | 500 rcs (10x1,25ml) | 320,00 | Add to cart |
Datasheet: dU-HOT Mastermix
Description:
2x SuperHot PCR Master Mix is an optimized ready-to-use PCR mixture of Taq DNA Polymerase, and antibodies to Taq DNA polymerase, PCR buffer, MgCl2 and dNTPs. dUTP in the Mixture provides the tool for carry-over protection of the sample with UDG (cat#111025) treatment. 2x PCR Master Mix contains all components for PCR, except DNA template and primers. The mixture was shown to be effective for Real Time PCR.
2x AB-SuperHot PCR Master Mix Composition
- Taq DNA Polymerase in reaction buffer: 0.1 units/µl
- Antibodies to Taq DNA polymerase, concentration adjusted for the effective inhibition of polymerase activity at 37oC
- 32 mM (NH4)2SO4
- 130 mM TrisHCl, pH 8.8 at 25oC
- 0,02% Tween-20
- 5,5 mM MgCl2
- dNTPs (dATP, dCTP, dGTP, dTTP): 0.4 mM of each
- dUTP - 0,6 mM
Pack size:
2x 1,25 ml - sufficient for 100 HotStart PCR reactions in 50 µl reaction volume
10x 1,2 5ml - sufficient for 500 HotStart PCR reactions in 50 µl reaction volume.
Supplied with the enzyme: 1 ml tube MgCl2 (100 mM)
Performance and purity tests:
Tested for the absence of endodeoxyribonucleases and exodeoxyribonucleases. The 2x SuperHot PCR 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 SuperHot PCR Master Mix with 1 µg of pUC19 DNA in 50 µl for 4 hours neither at 37°C nor at 70oC.
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 a PCR program.
